U.S. patent application number 10/691123 was filed with the patent office on 2004-10-21 for treatment of diabetes.
Invention is credited to Brand, Stephen J., Cruz, Antonio, Hew, Yin, Pastrak, Aleksandra.
Application Number | 20040209801 10/691123 |
Document ID | / |
Family ID | 32180693 |
Filed Date | 2004-10-21 |
United States Patent
Application |
20040209801 |
Kind Code |
A1 |
Brand, Stephen J. ; et
al. |
October 21, 2004 |
Treatment of diabetes
Abstract
Compositions and methods are provided for islet neogenesis
therapy comprising a member of a group of factors that complement a
gastrin/CCK receptor ligand, with formulations, devices and methods
for sustained release delivery and for local delivery to target
organs.
Inventors: |
Brand, Stephen J.; (Lincoln,
MA) ; Cruz, Antonio; (Toronto, CA) ; Pastrak,
Aleksandra; (Toronto, CA) ; Hew, Yin;
(Thornhill, CA) |
Correspondence
Address: |
MINTZ, LEVIN, COHN, FERRIS, GLOVSKY
AND POPEO, P.C.
ONE FINANCIAL CENTER
BOSTON
MA
02111
US
|
Family ID: |
32180693 |
Appl. No.: |
10/691123 |
Filed: |
October 22, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60420399 |
Oct 22, 2002 |
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60420187 |
Oct 22, 2002 |
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60428100 |
Nov 21, 2002 |
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60428562 |
Nov 22, 2002 |
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Current U.S.
Class: |
530/402 ;
514/11.3; 514/11.5; 514/11.7; 514/11.8; 514/12.3; 514/12.6;
514/20.3; 514/6.9; 514/7.2; 514/7.7; 514/8.1; 514/8.2; 514/8.5;
514/8.8; 514/8.9; 514/9.1; 514/9.2; 514/9.5; 514/9.6 |
Current CPC
Class: |
A61P 3/10 20180101; A61K
38/26 20130101; C07K 14/595 20130101; A61K 38/27 20130101 |
Class at
Publication: |
514/012 |
International
Class: |
A61K 038/17 |
Claims
What is claimed is:
1. A method for treating diabetes, the method comprising
administering to a mammal in need thereof a therapeutically
effective amount of a composition comprising a gastrin/CCK receptor
ligand and a factor for complementing gastrin for islet neogenesis
therapy (a FACGINT), provided that the FACGINT is not an EGF
receptor ligand.
2. A method according to claim 1, wherein the FACGINT is at least
one selected from the group of: a Glucagon-like peptide 1 receptor
ligand; a Glucagon-like peptide 2 receptor ligand; a gastric
inhibitory polypeptide (GIP) receptor ligand; a keratinocyte growth
factor (KGF) receptor ligand; a dipeptidyl peptidase IV inhibitor;
a REG protein receptor ligand; a Growth Hormone receptor ligand; a
Prolactin (PRL) receptor ligand; an Insulin-like Growth Factor
(IGF) receptor ligand; PTH-related protein (PTHrP) receptor ligand;
hepatocyte growth factor (HGF) receptor ligand; a bone
morphogenetic protein (BMP) receptor ligand, a transforming growth
factor-.beta. (TGF-.beta.) receptor ligand; a laminin receptor
ligand; vasoactive intestinal peptide (VIP) receptor ligand; a
fibroblast growth factor (FGF) receptor ligand; a keratinocyte
growth factor receptor ligand; a nerve growth factor (NGF) receptor
ligand; an islet neogenesis associated protein (INGAP) receptor
ligand; an Activin-A receptor ligand; a vascular endothelial growth
factor (VEGF) receptor ligand; an erythropoietin (EPO) receptor
ligand; a pituitary adenylate cyclase activating polypeptide
(PACAP) receptor ligand; a granulocyte colony stimulating factor
(G-CSF) receptor ligand; a granulocyte-macrophage colony
stimulating factor (GM-CSF); a platelet-derived growth factor
(PDGF) receptor ligand; and a Secretin receptor ligand.
3. A method according to claim 1, wherein the FACGINT comprises a
Glucagon 1-like peptide receptor ligand which is a GLP-1 or
exendin-4.
4. A method according to claim 2, wherein the FACGINT comprises a
Growth Hormone receptor ligand comprising a Growth Hormone.
5. A method for treating diabetes, the method comprising:
contacting ex vivo a plurality of cells with a composition
comprising at least one FACGINT and a gastrin/CCK receptor ligand,
provided that the FACGINT is not an EGF receptor ligand; and
administering the cells to a mammal in need thereof, thereby
treating the diabetes.
6. The method according to claim 5, wherein the cells are
autologous.
7. The method according to either of claims 1 or 5, wherein
administering or contacting is providing the composition in an
amount effective to increase the amount of insulin secreting cells
in the mammal.
8. The method according to either of claims 1 or 5, wherein the
composition is administered systemically.
9. The method according to either of claims 1 or 5, wherein the
amount of the FACGINT in the composition is substantially less than
the minimum effective dose of the FACGINT required to reduce blood
glucose in the diabetic mammal in the absence of a gastrin/CCK
receptor ligand
10. The method according to either of claims 1 or 5, further
comprising measuring a parameter selected from the group of: blood
glucose, serum glucose, blood glycosylated hemoglobin, pancreatic
.beta. cell mass, serum insulin, pancreatic insulin content, and
morphometrically determined .beta. cell mass.
11. The method according to claim 5, wherein the cells are
pancreatic ductal cells.
12. The method according to claim 1, further comprising measuring a
parameter selected from the group of: amount of insulin secreting
cells, glucose responsiveness of insulating secreting cells, amount
of proliferation of islet precursor cells, and amount of mature
insulin secreting cells.
13. A method for inducing pancreatic islet neogenesis in a mammal,
the method comprising administering to the mammal a composition
comprising a combination of a FACGINT and a gastrin/CCK receptor
ligand provided that the FACGINT is not an EGF receptor ligand, in
an amount sufficient to increase proliferation of islet precursor
cells in pancreatic tissue, thereby inducing pancreatic islet
neogenesis.
14. A method for inducing pancreatic islet neogenesis in a mammal,
the method comprising administering a composition comprising a
combination of a FACGINT and a gastrin/CCK receptor ligand wherein
the FACGINT is not an EGF receptor ligand, in an amount sufficient
to increase the number of pancreatic insulin secreting .beta. cells
in the mammal.
15. The method according to claim 5, further comprising prior to
the contacting step, culturing the cells ex vivo.
16. A composition comprising a gastrin/CCK receptor ligand and a
FACGINT, provided that the FACGINT is not an EGF receptor
ligand.
17. The composition according to claim 16 in a dosage effective for
inducing differentiation of an islet precursor cell into a mature
insulin secreting cell.
18. The composition according to claim 16 in a pharmaceutically
acceptable carrier.
19. A kit for treating or preventing diabetes, containing a
composition comprising a gastrin/CCK receptor ligand and a FACGINT,
a container, and instructions for use, provided that the FACGINT is
not an EGF receptor ligand.
20. The kit according to claim 19, wherein the composition further
comprises a pharmaceutically acceptable carrier.
21. A method for expanding and differentiating stem cells into
insulin secreting cells in a diabetic recipient of implanted cells,
comprising implanting the stem cells in the recipient, and
administering to the recipient a composition containing an
effective dose of each of a gastrin/CCK receptor ligand and at
least one FACGINT provided that the FACGINT is not an EGF receptor
ligand.
22. The method according to claim 21, wherein the cells are
obtained from a human or a porcine.
23. The method according to claim 21, wherein the implanted cells
are obtained from pancreatic islets, umbilical chords, embryos, or
stem cell lines.
24. A method according to any of claims 1, 5, 13, 14 and 21,
wherein the gastrin/CCK receptor ligand is gastrin.
25. A method according to claim 24, wherein the gastrin is
gastrin-17.
26. The method according to claim 24, wherein the gastrin/CCK
receptor ligand is human gastrin 1-17Leu15.
27. The method according to claim 23, wherein implanting the cells
in the recipient is using a route selected from: injecting directly
into an organ, and administering intravenously.
28. The method according to claim 23, wherein administering the
cells is delivering locally into an organ selected from the
pancreas, the kidney, and the liver.
29. The method according to claim 28, wherein delivering the cells
locally is a step selected from the group consisting of: endoscopic
retrograde cholangiopancreatography (ERCP); endoscopic
ultrasound-guided fine needle delivery (EUS-FNAD); injection into a
pancreatic artery; injection into a portal vein; intrapancreatic
injection; and injection into an hepatic artery.
30. The method according to claim 27, wherein injecting the cells
is delivering to the portal vein percutaneously or
transhepatically.
31. The method according to claim 23, further comprising prior to
the implanting step, treating the cells ex vivo with the
composition.
32. A method for reducing an amount of stem cells needed for
transplantation to treat human diabetes, the method comprising
administering to the recipient an effective dose of each of a
gastrin/CCK receptor ligand and a FACGINT provided that the FACGINT
is not an EGF receptor ligand, wherein the amount of cells needed
is reduced in comparison to an amount of cells needed in the
absence of administering the effective dose to an otherwise
identical recipient.
33. The method according to either of claims 1 and 2, further
comprising administering to the subject an agent for suppressing an
immune response.
34. The method according to claim 33, wherein the agent for
suppressing immune response is a drug.
35. The method according to claim 32, wherein the agent for
suppressing immune response is selected from at least one of the
group consisting of a rapamycin; a corticosteroid; an azathioprine;
mycophenolate mofetil; a cyclosporine; a cyclophosphamide; a
methotrexate; a 6-mercaptopurine; FK506; 15-deoxyspergualin; an FTY
720; a mitoxantrone; a 2-amino-1,3-propanediol; a
2-amino-2[2-(4-octylphenyl)ethyl]propane-1,3-d- iol hydrochloride;
a 6-(3-dimethyl-aminopropionyl) forskolin; and a
demethimmunomycin.
36. The method according to claim 32, wherein the agent for
suppressing immune response is a protein.
37. The method according to claim 36, wherein the protein comprises
an amino acid sequence of an antibody.
38. The method according to claim 37, wherein the agent for
suppressing immune response is selected from the group consisting
of at least one of: hul 124; BTI-322; allotrap-HLA-B270; OKT4A;
Enlimomab; ABX-CBL; OKT3; ATGAM; basiliximab; daclizumab;
thymoglobulin; ISAtx247; Medi-500; Medi-507; Alefacept; efalizumab;
infliximab; and an interferon.
39. The method according to claim 32, wherein the islet neogenesis
therapy composition and the agent for suppressing immune response
are administered sequentially.
40. The method according to claim 32, wherein at least one of the
islet neogenesis therapy composition and the agent for suppressing
immune response is administered systemically.
41. The method according to claim 40, wherein the islet neogenesis
therapy composition is administered as a bolus.
42. The method according to claim 32, wherein at least one of the
islet neogenesis therapy composition and the agent for suppressing
immune response is administered by a route selected from the group
consisting of intravenous, subcutaneous, intraperitoneal, and
intramuscular.
43. The method according to claim 32, wherein the agent for
suppressing immune response is administered orally.
44. The method according to claim 32, wherein the agent for
suppressing immune response is at least one selected from the group
of FK506, rapamycin, and daclizumab.
45. The method of according to either of claims 1 or 32, wherein
the subject is a human.
46. A kit for treatment of a diabetic subject, comprising an
immunosuppressive agent, an INT composition comprising a FACGINT
provided that the FACGINT is not an EGF receptor ligand, and a
container.
47. A pharmaceutical composition comprising a FACGINT provided that
the FACGINT is not an EGF receptor ligand and an agent for immune
suppression.
48. A pharmaceutical composition for sustained release of an
I.N.T..TM. therapeutic composition, the composition comprising: a
gastrin receptor ligand; and an EGF receptor ligand or a FACGINT;
wherein at least one of the gastrin receptor ligand, or the EGF
receptor ligand or FACGINT, is a sustained release formulation.
49. The composition according to either of claims 16 and 48,
further comprising an agent for immune suppression.
50. The composition according to claim 48, wherein the sustained
release formulation is selected from the group consisting of
pegylation and a multivesicular lipid-based liposome.
51. The composition according to claim 48, wherein the EGF receptor
ligand is selected from the group consisting of an EGF and a
TGFA.
52. A composition according to claim 48, wherein the FACGINT is at
least one selected from the group of: a Glucagon-like peptide 1
receptor ligand; a Glucagon-like peptide 2 receptor ligand; a
gastric inhibitory polypeptide (GIP) receptor ligand; a
keratinocyte growth factor (KGF) receptor ligand; a dipeptidyl
peptidase IV inhibitor; a REG protein receptor ligand; a Growth
Hormone receptor ligand; a Prolactin (PRL) receptor ligand; an
Insulin-like Growth Factor (IGF) receptor ligand; PTH-related
protein (PTHrP) receptor ligand; hepatocyte growth factor (HGF)
receptor ligand; a bone morphogenetic protein (BMP) receptor
ligand, a transforming growth factor-.beta. (TGF-.beta.) receptor
ligand; a laminin receptor ligand; vasoactive intestinal peptide
(VIP) receptor ligand; a fibroblast growth factor (FGF) receptor
ligand; a keratinocyte growth factor receptor ligand; a nerve
growth factor (NGF) receptor ligand; an islet neogenesis associated
protein (INGAP) receptor ligand; an Activin-A receptor ligand; a
vascular endothelial growth factor (VEGF) receptor ligand; an
erythropoietin (EPO) receptor ligand; a pituitary adenylate cyclase
activating polypeptide (PACAP) receptor ligand; a granulocyte
colony stimulating factor (G-CSF) receptor ligand; a
granulocyte-macrophage colony stimulating factor (GM-CSF); a
platelet-derived growth factor (PDGF) receptor ligand; and a
Secretin receptor ligand.
53. The composition according to claim 51, wherein the EGF receptor
ligand is a low molecular weight drug.
54. The composition according to claim 48, formulated for
parenteral administration.
55. The composition according to claim 48, formulated for oral
administration.
56. The composition according to claim 50, formulated for a route
of administration selected from the group consisting of
subcutaneous, intraperitoneal, intravenous, and intramuscular
injection.
57. The composition according to claim 48, wherein at least one of
the gastrin receptor ligand, or the EGF receptor ligand or the
FACGINT, is formulated for systemic administration.
58. The composition according to any of claims 16 and 48,
formulated for local delivery.
59. The composition according to claim 58, wherein local delivery
is targeted to the pancreas.
60. The composition according to claim 58, wherein local delivery
is selected from the group consisting of: endoscopic retrograde
cholangiopancreatography (ERCP); endoscopic ultrasound-guided fine
needle aspiration delivery (EUS-FNAD); injection into a pancreatic
artery; injection into a portal vein; intrapancreatic injection;
and injection into an hepatic artery.
61. The composition according to any of claims 16 and 48,
formulated for a route of administration selected from the group
consisting of transdermal and mucosal delivery.
62. The pharmaceutical composition according to claim 48,
formulated for delivery by a mechanical device.
63. The composition according to any of claims 16 and 48,
formulated for administration with a device selected from the group
of: a degradable implant; a transcutaneous patch; a catheter; an
implantable pump; a percutaneous pump; an infusion pump; and an
iontophoresis device.
64. The pharmaceutical composition according to claim 48,
formulated for a route of administration selected from the group
consisting of: subcutaneous, intraperitoneal, intravenous, and
intrapancreatic.
65. The pharmaceutical composition according to claim 64, wherein
the intravenous route is into a portal vein.
66. The pharmaceutical composition according to claim 62, wherein
the device is a pump.
67. The pharmaceutical composition according to claim 62, wherein
the administration is local.
68. The composition according to claim 48, wherein the
administration is local and is delivered by a route selected from
the group of: endoscopic retrograde cholangiopancreatography
(ERCP); endoscopic ultrasound-guided fine needle aspiration
delivery (EUS-FNAD); injection into a pancreatic artery; injection
into a portal vein; intrapancreatic injection; and injection into
an hepatic artery
69. The pharmaceutical composition according to claim 62, wherein
the administration is systemic.
70. The pharmaceutical composition according to claim 48, in an
effective dose.
71. A kit comprising at least one dose of a composition according
to claim 48.
72. A method of reducing frequency of treating a diabetic subject
with an I.N.T..TM. composition, the method comprising: preparing at
least one component of the composition as a sustained release
formulation; and administering the composition to the subject
according to a protocol having greater intervals between treatments
than for the composition not so formulated and otherwise
identical.
73. The method according to claim 72, wherein administering is
delivering by a route selected from: endoscopic retrograde
cholangiopancreatography (ERCP); endoscopic ultrasound-guided fine
needle aspiration delivery (EUS-FNAD; injection into a pancreatic
artery; injection into a portal vein; intrapancreatic injection;
and injection into an hepatic artery.
74. A method of enhancing efficacy of an I.N.T..TM. composition in
a diabetic subject, the method comprising: administering to the
subject an I.N.T..TM. composition having at least one component of
the composition formulated to produce a sustained release; and
comparing efficacy in treating the subject of an amount of the
composition administered to efficacy of a composition not having a
component so formulated and otherwise identical, such that the
efficacy of the I.N.T..TM. composition having a sustained release
formulated composition, as measured by a decrease in an amount of
the sustained release formulated agent required to reduce or
eliminate symptoms of diabetes in the subject, is enhanced.
75. The method of claim 74, wherein comparing efficacy is further
analyzing toxicity of the composition, such that fewer or milder
unwanted symptoms following administering the composition indicates
decreased toxicity in the composition having at least one component
formulated to produce a sustained release, compared to toxicity of
the I.N.T..TM. composition not having a component so formulated and
otherwise identical.
76. The method according to any of claims 73-75, wherein the
sustained release formulation of the component is selected from the
group consisting of pegylation and a multivesicular lipid-based
liposome.
77. The method according to any according to any of claims 73-76,
wherein the component having the sustained release formulation is
an EGF receptor ligand selected from the group consisting of an EGF
and a TGF.alpha..
78. The method according to any of claims 73-76, wherein the
FACGINT is at least one selected from the group of: a Glucagon-like
peptide 1 receptor ligand; a Glucagon-like peptide 2 receptor
ligand; a gastric inhibitory polypeptide (GIP) receptor ligand; a
keratinocyte growth factor (KGF) receptor ligand; a dipeptidyl
peptidase IV inhibitor; a REG protein receptor ligand; a Growth
Hormone receptor ligand; a Prolactin (PRL) receptor ligand ; an
Insulin-like Growth Factor (IGF) receptor ligand; PTH-related
protein (PTHrP) receptor ligand; hepatocyte growth factor (HGF)
receptor ligand; a bone morphogenetic protein (BMP) receptor
ligand, a transforming growth factor-.beta. (TGF-.beta.) receptor
ligand; a laminin receptor ligand; vasoactive intestinal peptide
(VIP) receptor ligand; a fibroblast growth factor (FGF) receptor
ligand; a keratinocyte growth factor receptor ligand; a nerve
growth factor (NGF) receptor ligand; an islet neogenesis associated
protein (INGAP) receptor ligand; an Activin-A receptor ligand; a
vascular endothelial growth factor (VEGF) receptor ligand; an
erythropoietin (EPO) receptor ligand; a pituitary adenylate cyclase
activating polypeptide (PACAP) receptor ligand; a granulocyte
colony stimulating factor (G-CSF) receptor ligand; a
granulocyte-macrophage colony stimulating factor (GM-CSF); a
platelet-derived growth factor (PDGF) receptor ligand; and a
Secretin receptor ligand.
79. The method according to claim 78, wherein the component is a
low molecular weight drug.
80. The method according to any of claims 73-76, wherein
administering is delivering by a route selected from the group
consisting of parenteral, oral, transdermal, subcutaneous, mucosal,
intraperitoneal, intravenous, intrapancreatic and
intramuscular.
81. The method according to claim 80, wherein administering
produces local distribution.
82. The method according to claim 82, further comprising
administering the composition in an effective dose.
83. The method according to claim 80, wherein prior to
administering, the composition is formulated for using a sustained
release device.
84. The method according to claim 83, wherein the device selected
from the group of: degradable implant; transcutaneous patch;
catheter; implantable pump; percutaneous pump; infusion pump; and
iontophoresis device.
85. The method according to claim 84, wherein the device is a
pump.
86. The method according to claim 80, wherein administering by the
intravenous route is injecting into a portal vein.
87. A method of reducing frequency of treating a diabetic subject,
the method comprising preparing a device for administering an
I.N.T..TM. composition to the subject by continuous release for a
prolonged period; providing the device to the subject; and
re-iterating treating the subject by replacing or refilling the
device.
88. A method according to claim 87, wherein the device is a
pump.
89. A method according to claim 88, wherein the pump is selected
from the group consisting of: a percutaneous pump; a flurorcarbon
propellant pump; an osmotic pump; a mini-osmotic pump; an
implantable pump; and an infusion pump.
90. A method according to claim 87, wherein the device is selected
from the group consisting of: a degradable implant; a
non-degradable implant; a mucoadhesive implant; a transcutaneous
patch; a catheter; and an iontophoresis device.
90. A method for expanding and differentiating stem cells into
insulin secreting cells in a diabetic recipient of the cells,
comprising: implanting the cells in the recipient; and
administering a sustained release composition comprising an
effective dose of each of: a gastrin/CCK receptor ligand; and a
FACGINT or an EGF receptor ligand, wherein the stem cells are
expanded and differentiated into insulin secreting cells in the
recipient.
91. A composition for treating diabetes comprising a Glucagon-like
peptide-1 (GLP-1) receptor ligand and a gastrin/CCK receptor
ligand.
92. The composition according to claim 91, wherein the GLP-1
receptor ligand is GLP-1.
93. A composition for treating diabetes comprising a Growth Hormone
(GH) receptor ligand and a gastrin/CCK receptor ligand.
94. The composition according to claim 93, wherein the GH is human
growth hormone (HGH).
96. A composition for treating diabetes comprising a prolactin (PL)
receptor ligand and a gastrin/CCK receptor ligand.
97. The composition according to claim 96, wherein the PL is human
PL.
98. The compositions according to any of claims 91-97, wherein the
gastrin is gastrin I having 17 amino acids with a Leu residue at
amino acid position 15.
99. The compositions according to any of claims 91-98, further
comprising an agent for immune suppression.
100. The compositions according to any of claim 91-99, further
formulated for sustained release.
101. A method of treating a diabetic subject comprising
administering to the subject a composition comprising a gastrin/CCK
receptor ligand and a Glucagon-like peptide-1 (GLP-1) receptor
ligand.
102. A method of treating a diabetic subject comprising
administering to the subject a composition comprising a gastrin/CCK
receptor ligand and a Growth Hormone (GH) receptor ligand.
103. A method of treating a diabetic subject comprising
administering to the subject a composition comprising a gastrin/CCK
receptor ligand and a prolactin (PL) receptor ligand.
104. The methods of any of claims 101-103 further comprising
administering an agent for immune suppression.
105. The methods of any of claims 101-104 further comprising
administering at least one of the receptor ligands or agents using
a sustained release device.
106. The methods of any of claims 101-104 further comprising
formulating at least one of the receptor ligands or agents for
sustained release.
107. The methods of any of claims 101-104 wherein the diabetic
subject has type I diabetes or type II diabetes.
108. A method for expanding a functional .beta. cell mass of
pancreatic islet transplants in a diabetic patient recipient of a
transplant of purified islets, the method comprising administering
to the mammal an effective dose of a gastrinCCK receptor ligand and
a FACGINT.
109. A method of treating human diabetes comprising transplanting a
pancreatic islet preparation into a diabetic patient; and
administering to the patient an effective dose of a gastrin/CCK
receptor ligand and a FACGINT.
110. A method according to claim 1, wherein the FACGINT comprises a
prolactin receptor ligand which is prolactin.
Description
RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 60/420,399 filed Oct. 22, 2002; U.S. Provisional
Application No. 60/420,187 filed Oct. 22, 2002; U.S. Provisional
Application No. 60/428,100 filed Nov. 21, 2002; and U.S.
Provisional Application No. 60/428,562 filed Nov. 22, 2002, each of
which is incorporated herein in its entirety. This application is
related to commonly owned U.S. application Ser. No. 10/000,840
filed Oct. 23, 2001, and to commonly owned U.S. application Ser.
No. 10/044,048 filed on Jan. 11, 2002, which claims priority to
U.S. provisional application 60/261,638, filed Jan. 12, 2001, and
to commonly owned U.S. application Ser. No. 10/426,998 filed Apr.
30, 2003, which claims priority to U.S. provisional application
60/377,180 filed Apr. 30, 2002, and to commonly owned U.S.
application Ser. No. 10/457,126 filed Jun. 9, 2003, which claims
priority to U.S. provisional application 60/387,032 filed Jun. 7,
2002.
FIELD OF THE INVENTION
[0002] The invention relates to methods and compositions for
treating a diabetic subject with islet neogenesis therapy and an
agent for immunosuppression, with formulations and methods for
local delivery and for sustained release of the compositions.
BACKGROUND OF THE INVENTION
[0003] The severe forms of the common disease Diabetes Mellitus
result from an absence or relative deficiency of insulin secretion
from the pancreatic .beta. cell. Consequently, diabetics are
dependent on exogenous insulin injections to prevent life
threatening complications of high blood glucose (hyperglycemia).
Unless patients adhere to a very demanding regime of glucose
testing and insulin treatment (intensive insulin treatment),
insulin treatment does not prevent chronic long-term complications
of organ damage caused by hyperglycemia. Intensive insulin
treatment increases risk of acute hypoglycemia due to insulin
overdosing, with acute and serious alterations of consciousness
state that can be fatal.
[0004] About one million people in the United States population
suffer from juvenile or type I diabetes, and about 30,000 new cases
arise each year. Further, an extremely large and rapidly increasing
number of patients have forms of type II diabetes (also called
adult onset or insulin-resistance diabetes), at a level of epidemic
proportions, a disease that can cause pancreatic exhaustion and
insulin insufficiency.
[0005] The abnormally high blood glucose (hyperglycemia) that
characterizes diabetes, if left untreated, results in a variety of
pathological conditions, for example, non-healing peripheral
vascular ulcers, retinal damage leading to blindness, and kidney
failure. Diabetes of both types I and II are treated with insulin
injection in response to blood glucose levels determined by patient
glucose self-monitoring, although not all type II patients progress
to requiring insulin therapy. Typically, multiple doses of insulin
are delivered by the patient per day. Severe pathological
consequences of diabetes are correlated with less rigorous control
of blood glucose level.
[0006] A potential treatment for diabetes would be to restore
.beta. cell function so that insulin release is dynamically
regulated in response to changes in blood glucose levels. This can
be achieved by pancreas transplantation, but this approach is
typically limited to diabetics requiring kidney transplants for
renal failure. Also, pancreas transplantation can require life-long
immunosuppression to prevent allogeneic graft rejection and
autoimmune destruction of the transplanted pancreas.
[0007] Recently, transplants of isolated human islet preparations
have successfully reversed insulin diabetes in human subjects for
prolonged periods. However, a large amount of donor islet cell
material is required for each recipient, and the supply of islet
cell material has not been sufficient to meet the demand.
SUMMARY
[0008] A feature of the invention is a method for treating a
diabetic condition, the method including administering to a mammal
a therapeutically effective amount of a composition comprising a
gastrin/CCK receptor ligand and a factor for complementing gastrin
for islet neogenesis therapy (a FACGINT), provided that the FACGINT
is not an EGF receptor ligand,
[0009] As the term is used herein, the FACGINT is at least one
selected from the group of: a Glucagon-like peptide 1 receptor
ligand; a Glucagon-like peptide 2 receptor ligand; a gastric
inhibitory polypeptide (GIP) receptor ligand; a keratinocyte growth
factor (KGF) receptor ligand; a dipeptidyl peptidase IV inhibitor;
a REG protein receptor ligand; a Growth Hormone receptor ligand; a
Prolactin (PRL) receptor ligand; an Insulin-like Growth Factor
(IGF) receptor ligand; PTH-related protein (PTHrP) receptor ligand;
hepatocyte growth factor (HGF) receptor ligand; a bone
morphogenetic protein (BMP) receptor ligand, a transforming growth
factor-.beta. (TGF-.beta.) receptor ligand; a laminin receptor
ligand; vasoactive intestinal peptide (VIP) receptor ligand; a
fibroblast growth factor (FGF) receptor ligand; a keratinocyte
growth factor receptor ligand; a nerve growth factor (NGF) receptor
ligand; an islet neogenesis associated protein (INGAP) receptor
ligand; an Activin-A receptor ligand; a vascular endothelial growth
factor (VEGF) receptor ligand; an erythropoietin (EPO) receptor
ligand; a pituitary adenylate cyclase activating polypeptide
(PACAP) receptor ligand; a granulocyte colony stimulating factor
(G-CSF) receptor ligand; a granulocyte-macrophage colony
stimulating factor (GM-CSF); a platelet-derived growth factor
(PDGF) receptor ligand and a Secretin receptor ligand.
[0010] As used herein, the term "diabetes" means any physiologic
indication of a short age of insulin, a production of antibodies
against insulin, or an excess of blood sugar. Diabetes is
exemplified but not limited to diabetes I, diabetes II, gestational
diabetes, and a pre-diabetic condition. As used herein, the term
"mammal" has the usual meaning of any member of Mammalia, and
includes humans.
[0011] In related embodiments, the FACGINT is a Glucagon-like
peptide 1 (GLP-1) receptor ligand; a Glucagon-like peptide 2
(GLP-2) receptor ligand; or a member of a Growth Hormone receptor
ligand family, and can be a Growth Hormone, such as Human Growth
Hormone (HGH), a Placental lactogen (PL), or a Prolactin (PRL), or
an exendin such as exendin-4.
[0012] Another feature of the invention provides a method for
treating diabetes, the method including contacting ex vivo a
plurality of cells with a composition having at least one of a
FACGINT and a gastrin/CCK receptor ligand, provided that the
FACGINT is not an EGF receptor ligand; and administering the cells
to a mammal in need thereof, thereby treatingg the diabetes. In one
embodiment of the method the cells are autologous, i.e., from the
subject. Alternatively, the cells are from another individual in
the same species, or even from another species. In the method using
cells ex vivo, the cells can be pancreatic ductal cells. Pancreatic
cells are a source of islet precursor cells. A further embodiment
of this method involves, prior to the implanting step, treating the
cells ex vivo with the composition. A related method further
includes, prior to the contacting step, culturing the cells ex
vivo.
[0013] Generally, the terms "administering" or "contacting" mean
that the user of the method is provided the composition in an
amount effective to increase the amount of insulin secreting cells
in the mammal.
[0014] Generally in these methods, the composition is administered
systemically. Further, the amount of the FACGINT in the composition
is substantially less than the minimum effective dose of the
FACGINT required to reduce blood glucose in the diabetic mammal in
the absence of a gastrin/CCK receptor ligand. The method can
further include measuring a parameter selected from the group of:
blood glucose, serum glucose, blood glycosylated hemoglobin,
pancreatic .beta. cell mass, serum insulin, pancreatic insulin
content, and morphometrically determined .beta. cell mass. In
general, one of skill in the art of diagnosis of diabetes would
measure a glucose level in blood or serum following a fast, i.e., a
period of no feeding of the subject or patient, of duration typical
of such a diagnosis. The standard measurement is an assay of
fasting blood glucose, or FBG.
[0015] The in vivo methods herein can further include measuring a
parameter selected from the group of: amount of insulin secreting
cells, glucose responsiveness of insulating secreting cells, amount
of proliferation of islet precursor cells, and amount of mature
insulin secreting cells, these measurements being made in a mammal
that is an experimental animal such as a genetically diabetic mouse
(NOD mouse) or a rodent in which diabetes has been induced (with
streptozoticin or STZ).
[0016] Another embodiment herein is a method for inducing
pancreatic islet neogenesis in a mammal which is administering to
the mammal a composition comprising a combination of a FACGINT and
a gastrin/CCK receptor ligand provided that the FACGINT is not an
EGF receptor ligand, in an amount sufficient to increase
proliferation of islet precursor cells in pancreatic tissue,
thereby inducing pancreatic islet neogenesis.
[0017] Yet another embodiment herein in a method for inducing
pancreatic islet neogenesis in a mammal, the method comprising
administering a composition comprising a combination of a FACGINT
and a gastrin/CCK receptor ligand wherein the FACGINT is not an EGF
receptor ligand, in an amount sufficient to increase the number of
pancreatic insulin secreting .beta. cells in the mammal.
[0018] Accordingly, an embodiment of the invention is a composition
comprising a gastrin/CCK receptor ligand and a FACGINT, provided
that the FACGINT is not an EGF receptor ligand. The composition in
some embodiments is provided in a dosage effective for inducing
proliferation of islet precursor cells into an increased amount of
mature insulin secreting cells. Similarly, the composition in some
embodiments is provided in a dosage effective for inducing
differentiation of an islet precursor cell into a mature insulin
secreting cell.
[0019] The composition can be provided in a pharmaceutically
acceptable carrier.
[0020] Also provided herein is a kit for treatingg diabetes,
containing a composition comprising a gastrin/CCK receptor ligand
and a FACGINT, a container, and instructions for use, provided that
the FACGINT is not an EGF receptor ligand. The bit composition can
be provided in one or more unit doses. The composition of the kit
further includes a pharmaceutically acceptable carrier.
[0021] Another feature of the invention herein is a method for
expanding and differentiating stem cells into insulin secreting
cells in a diabetic recipient of implanted cells, which includes
implanting the stem cells in the recipient, and administering to
the recipient a composition containing an effective dose of each of
a gastrin/CCK receptor ligand and a FACGINT provided that the
FACGINT is not an EGF receptor ligand. In this method, the cells
can be obtained for example from a human or a porcine. Further, the
implanted cells are obtained from pancreatic islets, umbilical
chords, embryos, or stem cell lines. Alternatively, a method for
expanding and differentiating stem cells in a diabetic recipient of
the cells into insulin secreting cells, is:implanting the cells in
the recipient; and administering a sustained release composition
comprising an effective dose of each of a gastrin/CCK receptor
ligand, and a FACGINT or an EGF receptor ligand.
[0022] In general according to these methods, the gastrin/CCK
receptor ligand is gastrin. In related embodiments, the gastrin is
gastrin-17, for example, the gastrin is human gastrin
1-17Leu15.
[0023] Further, according to these methods, implanting the cells in
the recipient is using a route such as injecting directly into an
organ, and administering intravenously. Injecting the cells is
delivering locally into an organ, for example, the pancreas, the
kidney, and the liver. Further, injecting the cells is delivering
to the portal vein percutaneously or transhepatically. In any of
these methods, a catheter can be inserted into a vein or artery
leading from or to the organ, using an imaging technology, such as
ultrasound or MRI during the procedure.
[0024] Delivering the cells locally can be chosen from several
technologies, including endoscopic retrograde
cholangiopancreatography (ERCP); endoscopic ultrasound-guided fine
needle delivery (EUS-FNAD); injection into a pancreatic artery;
injection into a portal vein; intrapancreatic injection; and
injection into an hepatic artery. In these technologies the user
can be guided by one of several imaging technologies including
ultrasound or MRI during the procedure.
[0025] Another feature of the invention provided is a method for
reducing an amount of stem cells needed for transplantation to
treat human diabetes, the method including administering to the
recipient an effective dose of each of a gastrin/CCK receptor
ligand and a FACGINT, the amount of cells being reduced in
comparison to implanting cells in the absence of administering the
effective dose to an otherwise identical recipient, provided that
the FACGINT is not an EGF receptor ligand.
[0026] In a related embodiment of the invention, the invention
provides a composition comprising a gastrin/CCK receptor ligand and
at least one FACGINT, provided that the FACGINT is not an EGF
receptor ligand. The composition is provided in a dosage that is
effective for inducing differentiation of an islet precursor cell
into a mature insulin secreting cells. The composition can further
be provided in a pharmaceutically acceptable carrier.
[0027] A kit is provided for treating diabetes, the kit containing
a composition comprising at least one FACGINT provided that the
FACGINT is not an EGF receptor ligand, and a gastrin/CCK receptor
ligand, a container, and instructions for use.
[0028] Any of the above methods can further include administering
to the subject an agent for suppressing an immune response. In any
of the above methods, the components of the combination can be
delivered simultaneously, for example, within one hour or within
one day, or can be delivered sequentially, for example, at an
interval of greater than one day, greater than two or more days,
greater than one week.
[0029] An exemplary agent for suppressing immune response is a
drug, for example, is at least one of the group consisting of a
rapamycin; a corticosteroid; an azathioprine; mycophenolate
mofetil; a cyclosporine; a cyclophosphamide; a methotrexate; a
6-mercaptopurine; FK506; 15-deoxyspergualin; an FTY 720; a
mitoxantrone; a 2-amino-1,3-propanediol- ; a
2-amino-2[2-(4-octylphenyl)ethyl]propane-1,3-diol hydrochloride; a
6-(3-dimethyl-aminopropionyl) forskolin; and a
demethimmunomycin.
[0030] Alternatively, an exemplary agent for suppressing immune
response is a protein, for example, the protein comprises an amino
acid sequence of an antibody. Accordingly, the agent for
suppressing immune response is at least one of: hul 124; BTI-322;
allotrap-HLA-B270; OKT4A; Enlimomab; ABX-CBL; OKT3; ATGAM;
basiliximab; daclizumab; thymoglobulin; ISAtx247; Medi-500;
Medi-507; Alefacept; efalizumab; infliximab; and an interferon. The
islet neogenesis therapy composition and the agent for suppressing
immune response are administered sequentially or can be
administered simultaneously.
[0031] In certain embodiments, at least one of the islet neogenesis
therapy composition and the agent for suppressing immune response
is administered systemically. For example, the islet neogenesis
therapy composition is administered as a bolus. Thus, at least one
of the islet neogenesis therapy composition and the agent for
suppressing immune response is administered by a route that is
intravenous, subcutaneous, intraperitoneal, or intramuscular.
Further, in certain embodiments, the agent for suppressing immune
response is administered orally. In general, the agent for
suppressing immune response is at least one of FK506, rapamycin,
and daclizumab. Further, according to embodiments of any of the
methods herein other than those calling for measurement of certain
experimental parameters, the subject can be a human.
[0032] Also featured herein is a kit for treatment of a diabetic
subject, comprising a container, an immunosuppressive agent, and an
INT composition comprising a FACGINT provided that the FACGINT is
not an EGF receptor ligand.
[0033] Also provided here is a pharmaceutical composition for
sustained release of an I.N.T..TM. therapeutic composition, the
composition comprising: a gastrin receptor ligand; and an EGF
receptor ligand or a FACGINT; wherein at least one of the gastrin
receptor ligand, or the EGF receptor ligand or FACGINT, is a
sustained release formulation. This composition can further
comprising an agent for immune suppression. Exemplary embodiments
of the sustained release formulation are pegylation of at least one
of the components of the composition, and formulation of at least
one component in a multivesicular lipid-based liposome. Examples of
the EGF receptor ligand is selected from the group consisting of an
EGF and a TGF.alpha.. Examples of the FACGINT is at least one
selected from the group of: a Glucagon-like peptide 1 receptor
ligand; a Glucagon-like peptide 2 receptor ligand; a gastric
inhibitory polypeptide (GIP) receptor ligand; a keratinocyte growth
factor (KGF) receptor ligand; a dipeptidyl peptidase IV inhibitor;
a REG protein receptor ligand; a Growth Hormone receptor ligand; a
Prolactin (PRL) receptor ligand ; an Insulin-like Growth Factor
(IGF) receptor ligand; PTH-related protein (PTHrP) receptor ligand;
hepatocyte growth factor (HGF) receptor ligand; a bone
morphogenetic protein (BMP) receptor ligand, a transforming growth
factor-.beta. (TGF-.beta.) receptor ligand; a laminin receptor
ligand; vasoactive intestinal peptide (VIP) receptor ligand; a
fibroblast growth factor (FGF) receptor ligand; a keratinocyte
growth factor receptor ligand; a nerve growth factor (NGF) receptor
ligand; an islet neogenesis associated protein (INGAP) receptor
ligand; an Activin-A receptor ligand; a vascular endothelial growth
factor (VEGF) receptor ligand; an erythropoietin (EPO) receptor
ligand; a pituitary adenylate cyclase activating polypeptide
(PACAP) receptor ligand; a granulocyte colony stimulating factor
(G-CSF) receptor ligand; a granulocyte-macrophage colony
stimulating factor (GM-CSF); a platelet-derived growth factor
(PDGF) receptor ligand; and a Secretin receptor ligand.
Alternatively, the EGF receptor ligand is a low molecular weight
drug. The composition can be formulated for parenteral
administration. Alternatively, the composition can be formulated
for oral administration. The composition can be formulated for a
route of administration selected from the group consisting of
subcutaneous, intraperitoneal, intravenous, and intramuscular
injection. In one embodiment, at least one of the gastrin receptor
ligand, or the EGF receptor ligand or the FACGINT, is formulated
for systemic administration.
[0034] Further, the above compositions can be formulated for local
delivery, for example, the local delivery is targeted to the
pancreas. Exemplary types of local delivery include endoscopic
retrograde cholangiopancreatography (ERCP); endoscopic
ultrasound-guided fine needle aspiration delivery (EUS-FNAD);
injection into a pancreatic artery; injection into a portal vein;
intrapancreatic injection; and injection into an hepatic artery.
These can be combined with imaging technologies such as ultrasound
and MRI, performed during the procedure.
[0035] The composition in some embodiments is formulated for a
route of administration selected from the group consisting of
transdermal and mucosal delivery. Any of these compositions can be
formulated for delivery by a mechanical device, in combination with
formulation for sustained release, or alternatively, use of the
mechanical device to deliver the formulation over a sustained
period of time. The device is, for example, a degradable implant; a
transcutaneous patch; a catheter; an implantable pump; a
percutaneous pump; an infusion pump; and an iontophoresis
device.
[0036] The above compositions and pharmaceutical compositions can
be formulated for a route of administration selected from the group
consisting of: subcutaneous, intraperitoneal, intravenous, and
intrapancreatic. For example, the intravenous route is into a
portal vein. A device can be used, and the device can be a pump.
With sustained release formulations as with some of the above
methods and compositions, the administration can be local. For
example, the local administration can be delivered by a route
selected from the group of: endoscopic retrograde
cholangiopancreatography (ERCP); endoscopic ultrasound-guided fine
needle aspiration delivery (EUS-FNAD); injection into a pancreatic
artery; injection into a portal vein; intrapancreatic injection;
and injection into an hepatic artery Alternatively with the
sustained release formulations and devices, the administration can
be systemic. The pharmaceutical composition can be provided in an
effective dose. Also featured herein is a kit comprising at least
one dose of a composition of any of the above sustained release
formulations.
[0037] Also featured is a method of reducing frequency of treating
a diabetic subject with an I.N.T..TM. composition, the method
including preparing at least one component of the composition as a
sustained release formulation; and administering the composition to
the subject according to a protocol having greater intervals
between treatments than for the composition not so formulated and
otherwise identical. Administering can be delivering by a route
selected from: endoscopic retrograde cholangiopancreatography
(ERCP); endoscopic ultrasound-guided fine needle aspiration
delivery (EUS-FNAD; injection into a pancreatic artery; injection
into a portal vein; intrapancreatic injection; or injection into an
hepatic artery. Imaging technology can be used to guide a catheter
in place during these procedures.
[0038] Also featured is method of enhancing efficacy of an
I.N.T..TM. composition in a diabetic subject, the method including:
administering to the subject an I.N.T..TM. composition having at
least one component of the composition formulated to produce a
sustained release; and comparing efficacy in treating the subject
of an amount of the composition administered to efficacy of a
composition not having a component so formulated and otherwise
identical, such that the efficacy of the I.N.T..TM. composition
having a sustained release formulated composition, as measured by a
decrease in an amount of the sustained release formulated agent
required to reduce or eliminate symptoms of diabetes in the
subject, is enhanced. In an embodiment of this methods, comparing
efficacy is further analyzing toxicity of the composition, such
that fewer or milder unwanted symptoms following administering the
composition indicates decreased toxicity in the composition having
at least one component formulated to produce a sustained release,
compared to toxicity of the I.N.T..TM. composition not having a
component so formulated and otherwise identical. In any of these
methods, exemplary sustained release formulations of the component
are pegylation and a multivesicular lipid-based liposome.
[0039] In any of these methods enhancing efficacy of an I.N.T..TM.
composition, the component having the sustained release formulation
is an EGF receptor ligand selected from the group consisting of an
EGF and a TGF.alpha.. Alternatively in embodiments of these
methods, the FACGINT is at least one selected from the group of: a
Glucagon-like peptide 1 receptor ligand; a Glucagon-like peptide 2
receptor ligand; a gastric inhibitory polypeptide (GIP) receptor
ligand; a keratinocyte growth factor (KGF) receptor ligand; a
dipeptidyl peptidase IV inhibitor; a REG protein receptor ligand; a
Growth Hormone receptor ligand; a Prolactin (PRL) receptor ligand ;
an Insulin-like Growth Factor (IGF) receptor ligand; PTH-related
protein (PTHrP) receptor ligand; hepatocyte growth factor (HGF)
receptor ligand; a bone morphogenetic protein (BMP) receptor
ligand, a transforming growth factor-.beta. (TGF-.beta.) receptor
ligand; a laminin receptor ligand; vasoactive intestinal peptide
(VIP) receptor ligand; a fibroblast growth factor (FGF) receptor
ligand; a keratinocyte growth factor receptor ligand; a nerve
growth factor (NGF) receptor ligand; an islet neogenesis associated
protein (INGAP) receptor ligand; an Activin-A receptor ligand; a
vascular endothelial growth factor (VEGF) receptor ligand; an
erythropoietin (EPO) receptor ligand; a pituitary adenylate cyclase
activating polypeptide (PACAP) receptor ligand; a granulocyte
colony stimulating factor (G-CSF) receptor ligand; a
granulocyte-macrophage colony stimulating factor (GM-CSF); a
platelet-derived growth factor (PDGF) receptor ligand; and a
Secretin receptor ligand. Even further as an alternative, a
component is a low molecular weight drug. In any of these methods,
administering is delivering by a route selected from the group
consisting of parenteral, oral, transdermal, subcutaneous, mucosal,
intraperitoneal, intravenous, intrapancreatic and
intramuscular.
[0040] For example, administering produces local distribution.
Further, the method includes administering the composition in an
effective dose. Even further, the method includes, prior to
administering, the composition is formulated for using a sustained
release device. For example, the device selected from the group of:
degradable implant; transcutaneous patch; catheter; implantable
pump; percutaneous pump; infusion pump; and iontophoresis device.
For example, the device is a pump. Administering in any of these
methods may be by the intravenous route is injecting into a portal
vein.
[0041] Also featured is a method of reducing frequency of treating
a diabetic subject, the method comprising preparing a device for
administering an I.N.T..TM. composition to the subject by
continuous release for a prolonged period; providing the device to
the subject; and re-iterating treating the subject by replacing or
refilling the device, for example, a device which is a pump. The
pump can be a percutaneous pump; a flurorcarbon propellant pump; an
osmotic pump; a mini-osmotic pump; an implantable pump; or an
infusion pump. Alternatively, the device is selected from the group
consisting of: a degradable implant; a non-degradable implant; a
mucoadhesive implant; a transcutaneous patch; a catheter; and an
iontophoresis device. An exemplary non-degradable implant is
Silastic. Other examples of degradable implants can be at least one
of the materials selected from the group of: starch; vinylstarch;
dipropyleneglycol diacrylate (DPGDA); tripropyleneglycol diacrylate
(TPGDA); pectin; cellulose acetate; cellulose propionate; cellulose
acetate butyrate; cellulose acetate propionate (CAP); hydroxypropyl
cellulose (HPC); hydroxypropyl cellulose/cellulose acetate
propionate (HPC/CAP); methyl methacrylate (MMA); butyl methacrylate
(BMA); hydroxymethyl methacrylate (HEMA); ethyl hexyl acrylate
(EHA); octadecyl methacrylate (ODMA); and ethyleneglycol
dimethacrylate (EGDMA).
[0042] Also featured is a method for expanding and differentiating
stem cells into insulin secreting cells in a diabetic recipient of
the cells, having steps of: implanting the cells in the recipient;
and administering a sustained release composition comprising an
effective dose of each of: a gastrin/CCK receptor ligand; and a
FACGINT or an EGF receptor ligand, wherein the stem cells are
expanded and differentiated into insulin secreting cells in the
recipient.
[0043] Also featured is a composition for treating diabetes
comprising a Glucagon-like peptide-1 (GLP-1) receptor ligand and a
gastrin/CCK receptor ligand. For example, the GLP-1 receptor ligand
is GLP-1. Also featured is a composition for treating diabetes
comprising a Growth Hormone (GH) receptor ligand and a gastrin/CCK
receptor ligand. For example, the GH is human growth hormone (HGH).
Also featured is a composition for treating diabetes comprising a
prolactin (PL) receptor ligand and a gastrin/CCK receptor ligand.
For example, the PL is human PL. In any of these compositions, the
exemplary gastrin is gastrin I having 17 amino acids with a Leu
residue at amino acid position 15. Any of these compositions can
further have an agent for immune suppression. Any of these
compositions can further be formulated for sustained release.
[0044] Also featured is a method of treating a diabetic subject,
which is administering to the subject a composition comprising a
gastrin/CCK receptor ligand and a Glucagon-like peptide-1 (GLP-1)
receptor ligand. Also featured is a method of treating a diabetic
subject which is administering to the subject a composition
comprising a gastrin /CCK receptor ligand and a Growth Hormone (GH)
receptor ligand. Also featured is a method of treating a diabetic
subject which is administering to the subject a composition
comprising a gastrin/CCK receptor ligand and a prolactin (PL)
receptor ligand. Any of these methods can include administering an
agent for immune suppression. Any of these methods can further
include administering at least one of the receptor ligands or
agents using a sustained release device. Any of these methods can
include further include formulating at least one of the receptor
ligands or agents for sustained release. Any of these methods can
be used with the diabetic subject who has type I diabetes or type
II diabetes.
[0045] Also featured is a method for expanding a functional .beta.
cell mass of pancreatic islet transplants in a diabetic patient
recipient of a transplant of purified islets, the method being
administering to the mammal an effective dose of a gastrinCCK
receptor ligand and a FACGINT.
[0046] Also featured is a method for method of treating human
diabetes comprising transplanting a pancreatic islet preparation
into a diabetic patient; and administering to the patient an
effective dose of a gastrin/CCK receptor ligand and a FACGINT.
Accordingly in this method, the FACGINT comprises a prolactin
receptor ligand which is prolactin.
DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS
[0047] Compositions for islet neogenesis therapy (I.N.T..TM.)
comprise a gastrin/CCK receptor ligand, and an EGF receptor ligand
or a Factor Complementing Gastrin Islet Neogenesis Therapy
(FACGINT).
[0048] Treatment of diabetes by administration with a combination
of a FACGINT and a gastrin/CCK receptor ligand, for example,
gastrin, gives a surprising level of enhancement of potency,
efficacy and utility over treatment with the any single component
alone. For this reason, the term FACGINT as used herein means
"complementary" to administration of gastrin. The term
"complementary" is not necessarily meant to imply a synergism
between gastrin and the FACGINT, rather the term means that in
comparison to administration of gastrin and the FACGINT,
administration of the combination is more efficacious for
remediating the diabetes at the doses used in the combination.
[0049] The term, "receptor ligand" as used herein in connection
with a receptor for a particular ligand shall mean any composition
or compound that binds to, interacts with, or stimulates that
receptor.
[0050] The term "FACGINT" includes a large variety of growth
factors and growth hormones, agents that modify one or more of the
factors hormones, and ligands and effectors for one or more
receptors involved in binding of these growth hormones and growth
factors as these terms are generally understood, exemplified but
not limited to: a PTH-related protein (PTHrP) receptor ligand such
as PTHrP (PTHrP; Garcia-Ocana, A. et al., 2001, J. clin. Endocrin.
Metab. 86: 984-988); a hepatocyte growth factor (HGF) receptor
ligand such as HGF (HGF; Nielsen, J. et al., 1999, J Mol Med 77:
62-66); a fibroblast growth factor (FGF) such as FGF, a
keratinocyte growth factor (KGF) receptor ligand such as KGF; a
nerve growth factor (NGF) receptor ligand such as NGF; a gastric
inhibitory polypeptide (GIP) receptor such as GIP; a transforming
growth factor beta (TGF.beta.) receptor ligand such as TGF.beta.
(U.S. patent application No. 2002/0072115 published Jun. 13, 2002),
a laminin receptor ligand such as laminin-1; an islet neogenesis
associated protein (INGAP) receptor ligand such as INGAP; a bone
morphogenetic factor (BMP) receptor ligand such as BMP-2; a
vasoactive intestinal peptide (VIP) receptor ligand such as VIP; a
glucagon-like peptide 1 receptor ligand such as GLP-1 and
exendin-4, glucagon-like peptide 2 (GLP-2) receptor ligand such as
GLP-2, and dipeptidyl peptidase IV inhibitors which indirectly
affect the levels of GLP-1 (Hughes, T. et al., 2002, Am Diabetes
Assoc Abstract 272-or) by inhibiting an enzyme involved in its
integrity; a REG receptor ligand such as REG protein; a Growth
hormone (GH) receptor ligand such a GH, a Prolactin (PRL) receptor
ligand such as PRL and placental lactogen (PL); an Insulin-like
growth factor (Type 1 and 2) receptor ligands such as IGF1 and
IGF-2; an Erythropoietin (EPO) receptor ligand such as EPO
(http://www.drinet.org/html/august.sub.--2002_.htm); a betacellulin
(also considered to be a member of the EGF family); an Activin-A
receptor ligand such as Activin-A; a vascular endothelial growth
factor (VEGF) receptor ligand such as VEGF; a bone morphogenesis
factor (BMP) receptor ligand such as BMP-2; a vasoactive intestinal
peptide (VIP) receptor ligand such as VIP; a vascular endothelial
growth factor (VEGF) receptor ligand such as VEGF; a pituitary
adenylate cyclase activating polypeptide (PACAP) receptor ligand
such as PACAP; a granulocyte colony stimulating factor (G-CSF)
receptor ligand such as G-CSF; a granulocyte-macrophage colony
stimulating factor (GM-CSF) receptor ligand such as GM-CSH; a
platelet-derived growth factor (PDGF) receptor ligand such as PDGF
and a Secretin receptor ligand such as secretin.
[0051] For any of the growth factors, enzymes, enzyme inhibitors,
peptide, protein and hormone compounds herein that are indicated to
be an exemplary FACGINT, all known analogues, variants, and
derivatives, whether naturally occurring or made by mutagenesis or
designed and synthesized shall be considered equivalent to that
FACGINT. Also considered among equivalents are conjugates, i.e.,
compositions derived by addition of one or more of a chemical
group, and mixtures thereof. Encoding genes may be altered by, for
example, oligonucleotide directed mutagenesis to produce FACGINT
analogues thereof, such as the human recombinant analogues.
Further, an identity or location of one or more than one amino acid
residue may be changed by targeted mutagenesis. The primary amino
acid sequence of the protein may be augmented by conjugates, as by
glycosylation, acylation, or by addition of any other supplementary
molecules, such as one or more of a lipid, a phosphate, and/or an
acetyl group. Further, individual amino acid residues in the chain
may be modified by oxidation, reduction, or other derivatization.
The FACGINT may be cleaved to obtain any fragments which retain
activity. An agonist, a prodrug or a metabolite of a FACGINT is
equivalent to that FACGINT. The whole polypeptide or protein or any
fragment can be fused with any other peptide or protein such as
immunoglobulins and other cytokines. Variants of FACGINTs that are
proteinaceous in nature can result from alternative splicing of a
primary transcript, from proteolytic cleavage or from other
post-translational modifications including dimerization,
polymerization, phosphorylation, glycosylation, sulfation and
deamidation. Conjugates may include, for example, a composition
comprising the FACGINT coupled to a non-naturally occurring polymer
comprising a polyalkylene glycol moiety. The term also encompasses
derivatives obtained by chemically modifying one or more amino acid
residues of the parent peptide, for instance by alkylation,
acylation, ester formation or amide formation. Further, agents that
induce synthesis of the FACGINT or mimic the action of the FACGINT
are contemplated as equivalent compounds. The singular form,
"FACGINT", may mean any one or more compounds from the exemplary
FACGINTs shown herein.
[0052] In several uses of the term herein, the term FACGINT is
specified not to include an EGF receptor ligand, as is made plain
in the context. However EGF receptor ligands such as EGF and TGF
are capable of complementing gastrin for remediation of diabetic
conditions and are therefore exemplary FACGINTs as defined herein,
and are included as components in other embodiments of
compositions, methods and kits herein. Certain embodiments of
I.N.T. therapy comprising administration of an EGF receptor ligand
in combination with a gastrin/CCK receptor ligand have been
described previously (U.S. Pat. Nos. 5,885,956 and 6,288,301),
references that do not use these agents in certain combinations and
formulations as shown herein.
[0053] The term, "pancreatic progenitor cell" or "beta cell
progenitor" or "islet precursor cell" as used herein is a precursor
cell capable of differentiating into a pancreatic beta islet cell,
which may or may not have the characteristic of a stem cell that is
the ability to reproduce itself in an unlimited manner. "Beta cell
neogenesis" or "islet neogenesis" means formation of new beta cells
by differentiation, which may or may not have the characteristic of
a stem cell that is the ability to reproduce itself in an unlimited
manner. To secrete insulin in a "glucose-responsive" manner means
to secrete insulin according to the glucose concentration in the
blood. In a physiologically normal mammal, beta cells of the islets
of Langerhans secrete insulin as blood glucose level is elevated,
i.e., are induced to secrete insulin in response to blood glucose
level.
[0054] A receptor "agonist" is any composition without limitation,
for example, a polypeptide growth factor or cytokine that is
endogenously found in the mammal, or a variant or portion thereof,
or an analog, or any peptidomimetic or low molecular weight drug,
that has the capability to bind to and activate the receptor of the
endogenously found polypeptide factor. For any of the growth
factors or cytokines described herein, an equivalent is considered
one that is substantially identical in amino acid sequence, for
example, shares 50% sequence identity, shares 60% sequence
identity, shares 70% sequence identity, or shares 80% sequence
identity with a naturally occurring peptide or protein as described
herein. In other embodiments, the agonist compositions herein
include an agonist inducing agent, which are envisioned to be
substances that, when given to an animal or provided to a cell,
organ or tissue in culture, is capable of increasing the amount of
that agonist produced by the animal, cell, organ or tissue. For
example, a prolactin release peptide stimulates the secretion of
prolactin. A receptor ligand includes within the scope of the
definition a receptor agonist, for the receptor for any particular
FACGINT, whether or not the agonist is structurally related to the
FACGINT.
[0055] The invention in one embodiment provides a method for
treating diabetic conditions such as diabetes mellitus by
administering a composition comprising both a gastrin/CCK receptor
ligand, e.g. gastrin, and a FACGINT, e.g. GLP-1, PRL or GH. Without
being limited by any particular mechanism, the gastrin/CCK receptor
ligand and the FACGINT are provided in an amount of each sufficient
to effect differentiation of pancreatic islet precursor cells to
mature insulin-secreting cells. Each of the FACGINT and the
gastrin/CCK receptor ligand in the composition can be administered
systemically or locally. Alternatively, one or both of the FACGINT
and the gastrin/CCK receptor ligand can be expressed in situ, by
cells that have been provided with a nucleic acid fusion construct
in an expression vector. The fusion construct typically includes a
preprogastrin peptide precursor coding sequence, and also a coding
sequence for a FACGINT.
[0056] Administration of a gastrin/CCK receptor ligand and an EGF
receptor ligand can achieve prolonged efficacy of islet cell
neogenesis, such that a therapeutic benefit is retained long after
cessation of treatment. See PCT application PCT/US02/00685 (WO
02/055152), published 18 Jul. 2002. The duration of the therapeutic
benefit is greater than the duration of the protocol for
administration of the composition.
[0057] Regenerative differentiation of residual pluripotent
pancreatic ductal cells into mature insulin-secreting cells is
obtained with the provided compositions and methods for treatment
of diabetes mellitus, particularly juvenile onset diabetes, and by
therapeutic administration of this combination of factors or
compositions which are provided for systemic administration, or for
in situ expression within the pancreas. An approach to .beta. cell
replacement that eliminates a requirement for cell transplantation
is stimulation of .beta. cell regeneration. Although early studies
suggested that a .beta. cell has limited capacity for regeneration,
it has been increasingly realized that the insulin secreting .beta.
cells of the pancreas comprise a dynamic cell population. The mass
of .beta. cells can expand through proliferation of existing .beta.
cells (.beta. cell replication). During pregnancy, prolactin,
growth hormone (Holstad, M. et al., J. Endocrinol. 163:229-234),
and placental lactogen (Nielsen, J. H., et al., J. Mol. Med.
77:62-66, 1999) stimulate the proliferation of .beta. cells to
increase .beta. cell mass. However, this expanded mass of cells
depends on continuing hormonal stimulation. After parturition, the
expanded .beta. cell mass decreases to non-pregnant levels, in
response to the decrease in prolactin and placental lactogen
(Logothetopoulos, J., (Logothetopoulos, J. (1972) in Handbook of
Physiology (Am. Physiol. Soc., Washington, D.C.), Section 7,
Chapter 3, pp67-76).
[0058] From this physiological information, an important aspect in
evaluating .beta. cell regeneration in response to administration
of a gastrin receptor ligand and either an EGF receptor ligand or a
FACGINT is whether an expanded .beta. cell mass can persist for a
significant time after the cessation of treatment with growth
factors. Use of a sustained release formulation, in combination
with the I.N.T. composition, with or without an agent for immune
suppression, can avoid a requirement for frequent visits to a
medical setting, or for self-medication.
[0059] The neogenesis of .beta. cells is measured as an increase in
both cell number and cell mass, resulting in an increase in plasma
insulin levels or pancreatic insulin content. Prolonged efficacy in
treatment of diabetes is a desired outcome of I.N.T.
[0060] An embodiment of the present invention provides improved
methods and compositions for use of a FACGINT administered with a
gastrin/CCK ligand to treat diabetes. The present invention in one
embodiment provides a combination of gastrin with a FACGINT to
achieve greater efficacy, potency, and utility than achieved with a
component alone, resulting in an improved therapeutic ratio for the
combination. Treatment with a combination of gastrin and a FACGINT
gave a reduction in blood glucose that was greater than observed
after treatment with a component alone, and the reduction in blood
glucose was sustained for a prolonged period after ceasing
treatment. The phrase, "a FACGINT" as used herein can also mean
"one or more FACGINTs" or "at least one FACGINT".
[0061] The .beta. cell stimulant Exendin (a GLP-1 analog) improved
glucose tolerance and increased .beta. cell mass in a partial
pancreatectomy model with mild diabetes (Xu, G. et al., Diabetes
48:2270-2276, 1999). However, a causal relationship between the
improved glucose tolerance and .beta. cell regeneration was not
conclusively established. GLP-1, as shown here to be an exemplary
FACGINT, when administered alone suppresses appetite and enhances
the clearance of glucose by reducing insulin resistance, a process
which can be independent of .beta. cell stimulation. Thus the
finding that plasma insulin levels were lower, not higher, in an
Exendin treated group, suggests that the observed improved glucose
tolerance was not a result of .beta. cell stimulation. Furthermore,
evaluation of the effect of Exendin on .beta. cell growth is
complicated by the pancreatectomy model of diabetes that was used
in these studies. An inflammation resulting from the surgical
ablation of the pancreas causes expression of growth factors that
act on islets such as gastrin and TGF.alpha., which by themselves
stimulate islet regeneration. Indeed, enhanced .beta. cell
regeneration has been reported after pancreatectomy alone
(Bonner-Weir, S., Diabetes 42: 1715-1720, 1993; Sharma, A., et al.,
Diabetes 48:507-513, 1999). Thus, it has not previously been clear
that Exendin could stimulate enhanced .beta. cell regeneration in
the absence of these pancreatectomy-induced growth factors.
[0062] The term, "diabetes" as used herein means any manifested
symptoms of diabetes in any mammal including experimental animal
models, and including human forms such as type I and type II
diabetes, early stage diabetes, and a pre-diabetic condition
characterized by mildly decreased insulin or mildly elevated blood
glucose levels. A "pre-diabetic condition" describes a mammal
suspected of having a diabetic or related condition, for example,
not formally diagnosed with diabetes, but demonstrating a symptom
in terms of insulin or glucose level, and susceptibilty to diabetes
or a related condition due to family history, genetic
predisposition, or obesity in the case of type II diabetes, or has
previously had diabetes or a related condition and is subject to
risk of recurrence.
[0063] As used herein, the term "gastrin/CCK receptor ligand"
encompasses any compound that binds to, interacts with or
stimulates the gastrin/CCK receptor. Examples of such gastrin/CCK
receptor ligands are given in U.S. Pat. No. 6,288,301 issued Sep.
11, 2001, and include various forms of gastrin, such as gastrin 34
(big gastrin), gastrin 17 (little gastrin), and gastrin 8 (mini
gastrin); various forms of cholecystokinin such as CCK 58, CCK 33,
CCK 22, CCK 12 and CCK 8; and other gastrin/CCK receptor ligands.
In general, gastrin/CCK receptor ligands share a carboxy terminal
amino acid sequence Trp-Met-Asp-Phe-amide. Also contemplated are
active analogs, fragments and other modifications of the above,
including both peptide and non-peptide agonists or partial agonists
of the gastrin/CCK receptor such as A71378 (Lin et al., Am. J.
Physiol. 258 (4 Pt 1): G648, 1990).
[0064] Small forms of gastrin such as gastrin 17 are economically
prepared by peptide synthesis, and synthetic peptides are
commercially available. Synthetic human gastrin 17 such as human
gastrin 17 having methionine or leucine at position 15 are also
available from Bachem AG, Bubendorf, Switzerland, and from
Researchplus. Gastrin/CCK receptor ligands include also active
analogs, fragments and other modifications of the above ligands,
which for example share amino acid sequence with an endogenous
mammalian gastrin, for example, share 60% sequence identity, or 70%
identity, or 80% identity. Such ligands also include compounds that
increase the secretion of endogenous gastrins, cholecystokinins or
similarly active peptides from sites of tissue storage. Examples of
these are the gastric releasing peptide, omeprazole which inhibits
gastric acid secretion, and soya bean trypsin inhibitor which
increases CCK stimulation.
[0065] The method for treating diabetes mellitus in an individual
in need thereof includes administering to the individual a
composition that provides both a gastrin/CCK receptor ligand and a
FACGINT. Without being limited by any particular mechanism, the
gastrin/CCK receptor ligand and the FACGINT are provided in doses
sufficient to effect differentiation of pancreatic islet precursor
cells to mature insulin-secreting cells.
[0066] The term "treating" or "ameliorating" as used herein means
reducing or eliminating one or more symptoms of a diabetes. A
method provided herein for treating diabetes comprises
administering, without being limited by the specific mechanism, a
differentiation regenerative amount of both a gastrin/CCK receptor
ligand and a FACGINT, to a diabetic mammal, to stimulate islet
neogenesis to increase the number of functional glucose responsive
insulin secreting .beta. cells in the pancreas. The method is
effective for diabetes generally, including Type I or juvenile
diabetes mellitus. The combination of gastrin and FACGINT would
result in a significant enhancement of the islet neogenesis
response over that observed with the individual components. An
exemplary gastrin/CCK receptor ligand is gastrin, and exemplary
FACGINTs are GLP-1, PRL and GH.
[0067] Another embodiment of the invention herein is a method
comprising treating explanted pancreatic tissue of a mammal with a
gastrin/CCK receptor ligand and FACGINT ex vivo, and introducing
the treated pancreatic tissue to the mammal. Again in this method,
an exemplary gastrin/CCK receptor ligand is gastrin, and exemplary
FACGINTs are GLP-1, PRL and GH.
[0068] In another embodiment, the invention provides a method for
gastrin/CCK receptor ligand stimulation, the method comprising
providing a chimeric insulin promoter-gastrin fusion gene to
pancreatic cells and expressing the gene. In yet another
embodiment, a method of FACGINT stimulation is provided, comprising
expressing a FACGINT gene that was trangenically introduced into a
mammal, for example, a gene encoding a FACGINT, for example, GLP-1,
PRL or GH. The gastrin/CCK receptor ligand gene can similarly be
provided trangenically, preferably a human preprogastrin peptide
precursor gene as shown in U.S. Pat. No. 5,885,956.
[0069] As used herein the term mammal shall include without
limitation any members of the Mammalia, such as a human, an ape, a
rodent such as a mouse or rat, a dog, a cat, an agriculturally
important animal or a protein pig, a goat, a sheep, a horse, or an
ape such as a gorilla or a chimpanzee. An individual mammal may be
non-diabetic, pre-diabetic, or diabetic, as specified herein.
[0070] Modes of systemic administration include, but are not
limited to, transdermal, intrathecal, intramuscular,
intraperitoneal, intravenous, subcutaneous, intranasal, and oral
routes. The compounds may be administered by any convenient route,
for example, by infusion or bolus injection, by absorption through
epithelial or mucocutaneous linings (e.g., oral mucosa, rectal,
vaginal, nasal, and intestinal mucosa, etc.), and may be
administered together with other biologically active agents. An
exemplary route of administration is systemic, for example, by
subcutaneous injection.
[0071] The term, "prolactin" as used herein means any polypeptide
which shares substantial sequence similarity with an endogenous
mammalian prolactin as this term is known in the art of protein
factors, for example, human prolactin, and which possesses the
activity of a prolactin. Endogenous human prolactin is a 199 amino
acid polypeptide produced by the pituitary gland. The term
encompasses prolactin analogs which are deletions, insertions, or
substitution mutants of endogenous prolactin, and retain the
activity, and includes prolactins from other species and naturally
occurring variants. The prolactin function includes a composition
having agonist activity for the prolactin receptor, as disclosed in
U.S. Pat. Nos. 6,333,031 (activating amino acid sequence) and
6,413,952 (metal complexed receptor ligand agonist), and G12ORhGH,
which is an analog of human growth hormone that acts as a prolactin
agonist (Mode et al., 1966, Endocrinol. 137(2): 447-454), and a
ligand for the prolactin receptor as described in U.S. Pat. Nos.
5,506,107 and 5,837,460. Also included are prolactin-related
protein, S179D, human prolactin and placental lactogens.
[0072] PRL, GH and PL are members of a family of polypeptide
hormones that share a structural, immunological and biological
functions (reviewed in, "Pancreatic Growth and Regeneration", Ed.
N. Sarvetnick, Ch. 1. Brejie, T. et al., 1997), and therefore
referred to herein as the PRL/GH/PL family. PRL and GH are secreted
by the anterior pituitary of vertebrate animals. PRL is involved in
a broad range of biological functions that include osmoregulation,
reproduction, lactation, and immunomodulation. GH is associated
with physiological processes related to growth and morphogenesis.
The related receptor ligands are referred to as "PRL/GH/PL"
receptor ligands. Classification of FACGINTs into various groups
based on structural similarity of the peptides and proteins,
functional similarity with respect to complementation of gastrin,
functional similarity with respect to binding of one or more
receptors, are each within the scope of various embodiments of the
invention. Prolactin receptor ligands include PRL and PL, and
growth hormone receptor ligands include GH.
[0073] As used herein, the term "GLP-1 receptor ligand" encompasses
any compound that binds to, interacts with or stimulates the GLP-1
receptor. Examples of GLP-1 receptor ligand include GLP-1 and
exendin-4. Glucagon-like peptide-1 is synthesized in intestinal
endocrine cells in molecular forms GLP- 1 (having residues
conventionally designated as positions 7-36) which is an amide, and
and similarly as GLP-1(7-37). Initial studies of GLP-1 biological
activity in utilized the full length N-terminal extended forms of
GLP-1 (1-37 and 1-36 which latter is an amide). The larger GLP-1
molecules were generally lacking biological activity. It was later
found that removal of the first six amino acids resulted in a
shorter version of the GLP-1 molecule having substantially enhanced
biological activity.
[0074] The majority of circulating biologically active GLP-1 is
found in the GLP-1(7-36)amide form, with lesser amounts of the
bioactive GLP-1(7-37) form also detectable. See Orskov, C. et al.,
Diabetes 1994, 43: 335-339. Both peptides show about the same
amount of biological function. GLP-1 is secreted from gut endocrine
cells in response to nutrient ingestion and plays multiple roles in
metabolic homeostasis following nutrient absorption. Regulation of
GLP-1 occurs by N-terminal degradation of the peptide by Dipeptidyl
Peptidase (DPP-IV) -mediated cleavage at the position 2 alanine
residue. For an overview, see DPP-IV. The biological activities of
GLP-1 include stimulation of glucose-dependent insulin secretion
and insulin biosynthesis, inhibition of glucagon secretion and
gastric emptying, and inhibition of food intake. GLP-1 appears to
have a number of additional effects in the GI tract and central
nervous system, as reviewed in Drucker, D., Endocrin 142: 521-527,
2001. Exemplary GLP-1 compositions include: BIM 51077 (GLP-1 analog
resistant to DPP-IV digestion, available from Beaufour Ipsen);
AC2592 (GLP-1, from Amylin, San Diego Calif.); ThGLP-1 (GLP-1,
modified amino acids and fatty acid attachment, from
Theratechnologies, Saint-Laurent, Quebec, Canada); CJC-1131 also
known as DACTM:GLP-1 (GLP-1 analog engineered for covalent coupling
to albumin, Conjuchem, Montreal, Quebec, Canada), LY315902 and
sustained release LY315902 (DDP-IV resistant GLP-1 analog from Eli
Lilly, Indianapolis, Ind.); a low molecular weight GLP-1 mimetic;
Albugon (albumin: GLP-1 fusion peptide from Human Genome Sciences,
Rockville, Md.); Liraglutide also known as NN2211 (long acting
GLP-1 derivative that is obtained by acylation of the GLP-1
molecule, which upon entering the bloodstream, is extensively bound
to albumin which protects it from degradation by DPP-IV and reduces
renal clearance, available from Novo Nordisk, Denmark; Elbrond et
al., Diabetes Care 2002 Aug. 25(8): 1398-404).
[0075] Exendin-4, an example of an exendin, is a novel peptide from
Heloderma suspectum (Gila monster) venom, having 53% homology with
GLP-1(7-36)amide. It functions as a long-acting potent agonist of
the glucagon-like peptide 1 (GLP-1) receptor, as it is resistant to
degradation by DDP-IV. Exendin-4 has properties similar to GLP-1,
and regulates gastric emptying, insulin secretion, food intake, and
slucagon secretion. Examples of exendin-4 include exenatide
(synthetic form also known as AC2993, Amylin); exenatide LAR
(longactinf form); ZP10 (modified exendin-4 having addition of six
lysine residues, Aventis/Zealand Pharma); and AP10 (long acting
formulation, Alkermes, Cambridge Mass.). Physiological studies
indicate that sustained expression of exendin-4 in transgenic
mammals does not perturb glucose homeostasis, cell mass or food
intake (Biaggio, L. et al. J Biol Chem 275: 34472-34477, 2000), so
that the physiological effects of exendin-4 are not completely
understood.
[0076] Dipeptidyl peptidase IV (DPP-IV) inhibitors refer to
compounds that inhibit activity of DPP-IV, a membrane-associated
peptidase of 766 amino acids that includes in its substrates GLP-1,
GLP-2 and GIP. DPP-IV-mediated inactivation of GLP-1 is a
determinant of GLP-1 bioactivity in vivo. Examples of DPP-IV
inhibitors include isoleucine thiazolidide, valine-purrolidide,
NVP-DPP738 (Novartis, Cambridge, Mass.), LAF237 (Novartis), P32/98
(Probiodrug AG, Halle, Germany), and P93/01 (Probiodrug).
[0077] As used herein, the term "EGF receptor ligand" encompasses
compounds that stimulate the EGF receptor such that when
gastrin/CCK receptors in the same or adjacent tissue or in the same
individual are also stimulated, neogenesis of insulin-producing
pancreatic islet cells is induced. Examples of such EGF receptor
ligands include full length EGF, which is EGF1-53, and further
include EGF1-48, EGF1-49, EGF1-52, and fragments and active analogs
thereof. Other examples of EGF receptor ligands are TGF.alpha.
forms that include 1-48, 1-47, 1-51, and amphiregulin and pox virus
growth factor as well as any EGF receptor ligands that demonstrate
the same synergistic activity with gastrin/CCK receptor ligands.
These include active analogs, fragments and modifications of the
above. See also, Carpenter and Wahl, Chapter 4, in Peptide Growth
Factors (Eds. Sporn and Roberts), Springer Verlag, 1990.
[0078] The group of compounds comprises the EGF receptor ligands
further includes "modified EGF", which includes variants of normal
or wild type EGF. Modifications have been shown to affect one or
more biological activity such as the rate of clearance of EGF. The
term includes peptides having an amino acid sequence substantially
similar to that of human EGF, for example, with one or a few amino
acid substitutions at various residue positions.
[0079] Recombinant EGF forms have been genetically engineered to
have alterations in structure and activities, for example, EGF
having a methionine at position 21 replaced by a leucine residue
has been described (U.S. Pat. No. 4,760,023). Recombinant human EGF
(hEGF) having 51 residues, i.e., lacking the two C-terminal
residues at positions 52 and 53 of HEGF, and having a neutral amino
acid substitution at position 51, retain EGF activity and are more
resistant to protease degradation during a microbial production
process, and following administration to a subject. A series of
nucleic acid molecules have been described that encode a family of
proteins that have significant similarity to EGF and TGF.alpha. (WO
00/29438). EGF muteins (mutated EGF) having histidine at residue 16
replaced with a neutral or acidic amino acid have been described
(WO 93/03757), such forms retaining activity at low values of pH.
Chemical analogues and fragments of EGF and TGF.alpha. retain
ability to bind various members of the EGF receptor family (U.S.
Pat. No. 4,686,283). Various modifications of EGF or TGF.alpha.
confer advantageous properties affecting one or more of recombinant
protein production, in vitro and in vivo stability, and in vivo
activity. A exemplary recombinant modified EGF receptor ligand used
in the Examples herein is a C-terminus deleted form of human EGF of
51 amino acids in length, having asparagine at position 51
(referred to herein as EGF51N), which retains substantially full
I.N.T..TM. activity, and has in vivo and/or in vitro stability that
is that is at least about as great or greater than normal or wild
type hEGF (S. Magil et al., published May 15, 2003 as
PCT/US02/33907, and incorporated by reference herein in its
entirety).
[0080] The term, "growth hormone" as used herein encompasses any
polypeptide that shares substantial amino acid sequence identity
with an endogenous mammalian growth hormone and possesses a
biological activity of a mammalian growth hormone. Human growth
hormone is a polypeptide containing 191 amino acids in a single
chain, and a molecular weight of about 22 kDal (Goeddel et al.,
1979, Nature 281: 544-548; Gray et al., 1985, Gene 39: 247-254).
The term encompasses analogs having deletions, insertions or
substitutions and growth hormones from other species and naturally
occurring variants. See Cunningham et al., 1989, Science 243:
1330-1336, and 1989, Science 244: 1081-1085; and WO 90/05185, and
U.S. Pat. No. 5,506,107.
[0081] The term, "erythropoietin" (EPO) as used herein is any
endogenous mammalian EPO or variant thereof, or EPO receptor
agonist, for example the EPO mimetic EMP1 (Johnson et al., 2000,
Nephr Dial Tranpl 15:1274-1277); or mimetics described (Wrighton et
al., 1996, Science 273:458-464; U.S. Pat. No. 5,773,569;
Kaushansky, 2001, Ann NY Acad Sci 938: 131-138); an antibody having
EPO receptor agonist activity (see for example, U.S. Pat. No.
5,885,574; WO 96/40231); and amino acid sequence disclosed in U.S.
Pat. Nos. 6,333,031, and 6,413,952.
[0082] The term, "PACAP" as used herein means an endogenously
produced PACAP or analog or variant thereof that shares substantial
amino acid identity or similarity, or has biological activity as a
PACAP receptor agonist such as maxadilan (Moro et al., 1997, J Biol
Chem 272:966-970. Useful PACAP variants include without limitation,
38 amino acid and 27 amino acid variants as disclosed in U.S. Pat.
Nos. 5,128,242; 5,198,542; 5,208,320; and 6,242,563).
[0083] Pharmaceutical Compositions
[0084] The present invention in various embodiments provides
pharmaceutical compositions comprising a therapeutically effective
amount of a combination of a FACGINT, and a gastrin/CCK receptor
ligand. All of the pharmaceutical compositions described herein can
be formulated with or without an agent for immune suppression, and
with or without components or devices for sustained release, for
delivery locally or systemically. A pharmaceutically acceptable
carrier or excipient can be added. Such a carrier includes but is
not limited to saline, buffered saline, dextrose, water, glycerol,
ethanol, and combinations thereof. The formulation should suit the
mode of administration. An "effective amount" as the term is used
herein is an amount of a therapeutic agent or combination of agents
sufficient to achieve a recognized medical endpoint, in this case,
remediation of a symptom of diabetes. The effective amount can be
determined empirically by a skilled artisan according to
established methods of measurement of relevant parameters, as
described herein.
[0085] The compositions herein can further comprise wetting or
emulsifying agents, or pH buffering agents. The composition can be
a liquid solution, suspension, emulsion, tablet, pill, capsule,
sustained release formulation, or powder. The compositions can be
formulated as a suppository, with traditional binders and carriers
such as triglycerides. Oral formulation can include standard
carriers such as pharmaceutical grades of mannitol, lactose,
starch, magnesium stearate, sodium saccharine, cellulose, magnesium
carbonate, etc. Various delivery systems are known and can be used
to administer a composition of the invention, e.g., encapsulation
in liposomes, microparticles, microcapsules and the like.
[0086] In an exemplary embodiment, a composition herein is
formulated in accordance with routine procedures as a
pharmaceutical composition adapted, for example, for subcutaneous
administration to human beings. Typically, compositions for
subcutaneous administration are solutions in sterile isotonic
aqueous buffer. Where necessary, the composition may also include a
solubilizing agent and a local anesthetic to ameliorate pain at the
site of the injection. Generally, the ingredients are provided
either separately or mixed together in unit dosage form, for
example, as a dry, lyophilized powder or water-free concentrate in
a hermetically sealed container such as an ampoule or sachette, for
example, indicating the quantity of active agent. Where the
composition is to be administered by infusion, it can be dispensed
with an infusion bottle containing sterile pharmaceutical grade
water, buffer, or saline. Where the composition is administered by
injection, an ampoule of sterile water or saline for injection can
be provided so that the ingredients may be mixed prior to
administration. The compositions herein can in various components
thereof be formulated as suppositories, which contain active
ingredient in the range of about 0.5% to about 10% by weight; oral
formulations preferably contain about 10% to about 95% active
ingredient by weight.
[0087] A daily dose is administered as a single dose, or is divided
into a plurality of smaller fractional doses, to be administered
several times during the day.
[0088] The compositions of the invention can be formulated as
neutral or salt forms. Pharmaceutically acceptable salts include
those formed with free amino groups such as those derived from
hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and
those formed with free carboxyl groups such as those derived from
sodium, potassium, ammonium, calcium, ferric hydroxides,
isopropylamine, triethylamine, 2-ethylamino ethanol, histidine,
procaine, etc.
[0089] The amount of the therapeutic of the invention which will be
effective in the treatment of a particular disorder or condition
will depend on the nature of the disorder or condition, and can be
determined by standard clinical techniques. The precise dose to be
employed in the formulation will also depend on the route of
administration, and the seriousness of the disease or disorder, and
should be decided according to the judgment of the practitioner and
each patient's circumstances. Routine determinations of blood
levels of insulin or C peptide, and of fasting levels of glucose or
glucose challenges, are determined by one of ordinary skill in the
art. Effective doses may be extrapolated from dose-response curves
derived from in vitro or animal model test systems, by one of
ordinary skill in the art of pharmacology. Suitable dosage ranges
for administration are generally about 0.01 micrograms to about
10,000 micrograms of each active I.N.T..TM. compound per kilogram
body weight per day, for example, about 0.01 micrograms to about 1
microgram/kg, about 0.1 micrograms/kg to about 10 micrograms/kg,
about 1 microgram/kg to about 500 micrograms/kg, or about 10
micrograms/kg to about 10 mg/kg of body weight per day. Suitable
dosage ranges for administration are thus generally about 0.01
micrograms/kg body weight/day to about 10 mg/kg body
weight/day.
[0090] The invention in other embodiments provides a pharmaceutical
pack or kit comprising one or more containers filled with one or
more of the ingredients of the pharmaceutical compositions of the
invention. In such a pack or kit can be found a container having a
unit dosage of each or both of a gastrin/CCK receptor ligand and/or
one or more of a FACGINT, or an EGF receptor ligand, and one or
more of an immunosuppressive agent. One or more of these components
of the pack or kit can be formulated for sustained release or for
insertion as a refill into a sustained release device, or can be
formulated for local delivery. Associated with such container(s)
can be various written materials such as instructions for use, or a
notice in the form prescribed by a governmental agency regulating
the manufacture, use or sale of pharmaceuticals or biological
products, which notice reflects approval by the agency of
manufacture, use or sale for human administration. In some
embodiments the kit or pack may be associated with software
embedded in a computer readible format.
[0091] The invention in one aspect features islet neogenesis
therapy (I.N.T..TM.) compositions and methods, for example, a
gastrin and a FACGINT, in combination with immunosuppressive
agents, to stimulate the growth of new .beta. cells in vivo,
increasing islet mass, and result in improved glucose tolerance in
diabetic subjects, for example, in diabetic humans and in
animals.
[0092] The gastrin/CCK receptor ligand and the EGF receptor ligand
or FACGINT, can be administered in a single combined dose, or
administered separately in any order. An "effective combined dose"
of these compositions is a dose that produces a decrease in fasting
blood glucse, or an increase in amount of insulin secreting cells,
or an increase in insulin blood level, or an increase in
.beta.-cell mass. The gastrin/CCK receptor ligand is, in one
embodiment, human gastrin of length 17 amino acid residues, the
residue at position 15 being leucine (1-17Leu15, referred to herein
as gastrin17leu15); further, the EGF receptor ligand is human
EGF51N (S. Magil et al., published May 12, 2003 as PCT/US02/33907,
and incorporated by reference herein in its entirety). The
effective dose can contain a ratio of gastrin/CCK receptor ligand
to EGF receptor ligand that is greater than 1, for example, the
effective dose contains a ratio of gastrin/CCK receptor ligand to
EGF receptor ligand greater than 10. A convenient route of
administering the dose is with a systemic injection, for example, a
subcutaneous bolus.
[0093] In a further embodiment, the recipient subject is
administered an agent that suppresses the immune system. For
example, the agent is a low molecular weight organic chemical, for
example, is at least one of Tacrolimus, Sirolimus, cyclosporine,
and cortisone and other drugs as shown in Table 1. In an
alternative embodiment, the agent is an antibody, for example, the
antibody is anti-CD11 a and other antibodies also shown in Table 1.
In yet another alternative, the immunosuppressive agent can be an
antibody that is elaborated by the subject following an
immunization schedule, for example, against GFAP or against
S100.beta.. The subject can be diabetic, for example, the subject
is a non-obese diabetic mouse (the NOD mouse) or a
streptozoticin-treated mouse. The subject can be a human, for
example a diabetic patient having type I or type II diabetes, or
having a pre-diabetic condition, or having gestational diabetes, or
having had diabetes in the past, for example, having had
gestational diabetes in a past pregnancy.
[0094] Further, evaluating the size and function of newly developed
.beta. insulin secreting cells or islets is measuring a standard
physiological or diagnostic parameter including: islet .beta. cell
mass, islet .beta. cell number, islet .beta. cell percent, blood
glucose, serum glucose, blood glycosylated hemoglobin, pancreatic
.beta. cell mass, pancreatic .beta. cell number, fasting plasma C
peptide content, serum insulin, and pancreatic insulin content.
[0095] As diabetes is in certain cases an autoimmune disease, an
embodiment of I.N.T..TM. is systemic administration of
therapeutically effective doses of, for example, the ligands of
receptors for each of EGF and gastrin/CCK or each of a FACGINT and
gastrin/CCK, such as a combination of a gastrin and an EGF or a
combination of a FACGINT and gastrin/CCK, to subjects or patients
who are also treated with one or more agents that suppress the
immune system, i.e., immunosuppressive agents.
[0096] A number of different endpoints can be used to determine
whether gastrin and EGF or FACGINT treatment, or treatment with the
combination of gastrin and EGF or FACGINT and an immunosuppressive
agent, improves the diabetes, for example, increases the functional
mass of .beta. cells in the islet transplants. These include
measurement of enhanced plasma levels of circulating human C
peptide and human insulin, after injecting mice with .beta. cell
stimulants such as glucose or arginine; a response to gastrin/EGF
treatment demonstrated by increased human insulin immunoreactivity
or mRNA levels extracted from the islet transplants; and increased
number of .beta. cells, determined by morphometric measurement of
islets in treated animals.
[0097] The term "transplanting" as used herein means introducing a
cellular, tissue or organ composition into the body of a mammal by
any method established in the art, or as indicated herein. The
composition is a "transplant", and the mammal is the recipient. The
transplant and recipient may be syngeneic, allogenic, or
xenogeneic. The term, "autologous" as used herein means that the
transplant is derived from the cells, tissues or organs of the
recipient.
[0098] Enhanced .beta. cell function of human islets can also be
demonstrated by reversal of the hyperglycemia in recipient mice
with streptozotocin induced or genetic (using a strain of mice
known as non-obese diabetic or NOD) diabetes. Enhanced .beta. cell
function after treatment of the diabetic recipient subject with
gastrin, with EGF or FACGINT and with one or more immunosuppressive
agents is demonstrated by improved survival upon withdrawal of
insulin, and by correcting hyperglycemia as indicated by fasting
blood glucose level. Further, increases in both pancreatic insulin
and plasma C-peptide are observed.
1TABLE 1 Exemplary agents for immune suppression, and commercial
sources Names Company Nature 2-amino-1,3-propanediol Novartis Used
for preventing or derivatives treating chronic rejection in a
patient receiving an organ or tissue allo-or xeno- transplant
2-amino-2[2-(4- Yoshitomi Immunosuppression, from
octylphenyl)ethyl]propane- Pharmaceutical accelerated lymphocyte
1,3-diol hydrochloride Industries, Ltd homing
40-O-(2-hydroxyethyl)- Novartis Sirolimus (rapamycin) rapamycin,
SDZ-RAD, Pharmaceuticals derivative, used for acute Everolimus
(Certican .RTM.) kidney rejection; reduces rejection and graft
vasculopathy following heart transplantation by inhibiting cell
proliferation 6-(3-dimethyl- Matsumori Akia Immunosuppressing
action aminopropionyl) forskolin Nippon Kayaju Co useful also for
treating Ltd autoimmune disease 6-mercaptopurine Glaxo SmithKline
Used to treat Crohn's (Purinethol .RTM., 6-MP) disease,
inflammatory bowel disease and for organ transplant therapy ABX-CBL
(CBL-1) Abgenix Mouse monoclonal AB targeted against human T- cell,
B cells, NK cells and monocytes, fortreatment of steroid-resistant
graft vs host diseases, potential use in treatment of inflammatory
and autoimmune disorders Alefacept (human LFA-3 University of Utah-
Knocks out causative IgG1 fusion protein, Dermatology memory
T-lymphocytes; AMEVIVE .RTM.) Dept/BIOGEN Used to treat psoriasis,
a T- cell mediated inflammatory disorder HLA-B2702 peptide SangStat
Medical Human peptide, blocks (Allotrap .RTM.) action of NK cells
and T- cell mediated toxicities, used for prevention of first
kidney allograft rejection Antisense ICAM-1 ISIS-Boehringer Mouse
monoclonal AB inhibitor (ISIS 2302), Ingleheim blocks white blood
cell Enlimomab, BIRR1, adhesion to T-cell surface Alicaforsen)
molecule (ICAM-1r); treatment of kidney transplant rejection
Azathioprine (Imuran .RTM., Generic, Glaxo Treatment of rheumatoid
Azasan .RTM.) SmithKline, arthritis and prevention of Prometheus
kidney transplant rejection, Laboratories, and other autoimmune or
aaiPharma inflammatory disorders such as inflammatory bowel disease
BTI-322 MedImmune Mouse derived monoclonal AB targeted to CD2
receptor; used for prevention of first-time kidney rejection, and
treatment of resistant rejection Cladribine (Leustatin .RTM.)
Boehringer Antimetabolite and Ingleheim immunosuppressive agent
that is relatively selective for lymphocytes; used to treat
lymphoid malignancies, e.g., hairy- cell leukemia. Cyclophosphamide
(CTX, Generic Immunosuppressant t for Neosar .RTM., Cytoxan .RTM.,
treatment of arthritis and Procytox .RTM.) other auto-immune
disorders and cancers Cyclosporine (cyclosporin Novartis 11 amino
acid cyclic A, cyclosporin) peptide; blocks helper T- (Sandimmune
.RTM., Neoral .RTM., cell, immunosuppressant SangCya .RTM.) used in
organ transplant therapy and other immune diseases
Demethimmunomycin" (L- Merck & Co Treatment of autoimmune
683,742: also described as diseases, infectious diseases
31-desmethoxy-31- and/or prevention of organ hydroxy-L-683,590)
transplant rejections Dexamethasone Generic An adrenocorticoid,
(Decadron, Dexone, effective Dexasone) immunosuppressant in various
disorders Docosahexaenoic acid Not Immunosuppressant by that (DHA)
applicable lowers the proportion of T cells expressing CD4 or CD8,
blocks antigen recognition process; Taku et al., Journal of
Agricultural and Food Chemistry, 2000; 48(4): 1047 FTY720 (oral
myriocin Novartis Alters lymphocyte derivative) Pharmaceuticals
infiltration into grafted tissues; used for prevention of organ
rejection in kidney transplants Glatiramer acetate (copolymer-1,
Teva Synthetic peptide Copaxone .RTM.) Pharmaceuticals copolymer;
decoy that mimics structure of myelin so immune cells bind Copaxone
instead of myelin; for multiple sclerosis Glial fibrillary acidic
CalBiochem; Synx Possesses protein (GFAP) Pharma immunosuppressive
activities in diabetic animal models; Winer et al., Nature Medicine
9: 198 (2003) Gusperimus, (15- Bristol Myers-Squibb Intravenous
deoxyspergualin (Spanidin .RTM.) immunosuppressant; suppresses
production of cytotoxic T-cells, neutrophils and macrophages hu1124
(anti-CD11a) XOMA Humanized monoclonal antibody; targets CD11a
receptor on surface of T cells to selectively inhibit immune system
rejection of transplanted organs Infliximab (Remicade .RTM.)
Centocor (affiliate of Monoclonal AB, binds and Johnson and
inactivates human TNF- Johnson) alpha and; used to treat Crohn's
disease and rheumatoid arthritis Interferon Various companies
Immunomodulatory including Serono, properties Biogen etc ISAtx247
Isotechnika Used to treat autoimmune diseases such as rheumatoid
arthritis and psoriasis isotretinoin Immunosuppressant, reduces
ability of T cells to proliferate in response to immune challenge.
Vergelli et al., Immunopharmacology, 1997, 31: 191. Medi-500
(T10B9) MedImmune Intravenous monoclonal AB that targets human
T-cells; treats acute kidney rejection and graft-vs-host disease
Medi-507 MedImmune/Bio- Intravenous humanized AB Transplant
directed against CD2 T-cell; used to treat corticosteroid-
resistant graft vs host disease and prevention of kidney rejection
Methotrexate Wyeth Lederle, Antimetabolite used to treat
(Rheumatrex .RTM., Generic Crohn's disease, severe Amethopterin,
Trexall .RTM.) psoriasis, and adult rheumatoid arthritis (and as an
anti-cancer drug) Mitoxantrone Immunex Antiproliferative effect on
(Novantrone .RTM.) cellular immune system including T-cells,
B-cells and macrophages; used to treat hormone-refractory prostate
cancer, acute myelogenous leukemia and multiple sclerosis
mycophenolate mofetil Roche Proliferation of T and B (CellCept
.RTM.) lymphocytes by blocking the synthesis of purine nucleotides;
used in organ transplant therapy and inflammatory bowel disease
OKT4A R. W. Johnson Mouse monoclonal AB Pharmaceutical targeted
against human Research Institute CD4 T cell; used for prevention of
kidney transplant rejection when used in combination with other
immunosuppressant drugs Muromonab-CD3 R. W. Johnson Monoclonal AB
that binds (Orthoclone OKT3 .RTM.)() Pharmaceutical to receptor
sites on T-cells, Research Institute preventing activation by
transplanted tissue Prednisolone (Deltasone .RTM., Corticosteroid,
suppresses Oraone .RTM.) inflammation associated with transplant
rejection basiliximab (Simulect .RTM.) Novartis Monoclonal AB that
binds Pharmaceuticals to receptor sites on T-cells, preventing
activation by transplanted tissue (renal transplant) S100.beta.
glial protein Possesses immunosuppressive activities in diabetic
animal models Sirolimus, Rapamycin Wyeth-Ayerst Immunosuppressant
and (Rapamune .RTM.) Laboratories potent inhibitor of cytokine
(e.g.IL-2)-dependent T-cell proliferation (kidney transplant)
Tacrolimus (Prograf; FK- Fujisawa Interferes with IL-2 TCR 506)
communication Antithymocyte SangStat Medical Anti-human thymocyte
immunoglobulin Corporation, immunoglobulin; used in (ATGAM,
Thymoglobulin .RTM.) Pharmacia and reversal of acute kidney Upjohn
transplant rejection and will likely be used off-label for
transplant induction therapy efalizumab (Xanelim .RTM.) XOMA T-cell
modulator that target T-cells through interactions with adhesion
molecules on endothelial cell surface, target migration of T-cells
into the skin and target activation of T-cells; Used to treat
Psoriasis Daclizumab (Zenapax .RTM.), Protein Design Monoclonal AB
inhibits HAT (Humanized Anti- Laboratories/Roche binding of IL-2 to
IL-2 Tac), SMART anti-Tac, receptor by binding to IL-2 anti-CD25,
and humanized receptor; suppresses T cell anti-IL2-receptor
activity against allografts (renal transplant)
[0099] As used herein, the term "immunosuppressant" or "agent for
immune suppression" means any agent that suppresses immune
response. Exemplary immunosuppressant agents are shown in Table 1,
and any derivatives of those agents or functional equivalents are
considered appropriate for embodiments of the invention as
described herein and in the claims.
[0100] As used herein, a dosing schedule refers to a protocol for
administering any of the compositions provided herein, for example,
the components that make up the I.N.T..TM. composition or the
combination of a FACGINT and a gastrin/CCK receptor ligand, and one
or more of an immunosuppressive agent, each in an effective dose,
administered simultaneously or within a particular interval of each
other, for example, within one day of each other, or as a combined
preparation, or separately, and includes the amount of the
composition delivered per unit time such as per day, and the
duration or period of time over which each composition is
administered.
[0101] Most insulin dependent diabetic patients require insulin
injection at least on a daily basis. Multiple doses per day of
insulin are required under certain circumstances of illness or diet
for management of diabetes, and the insulin administration is
indicated by results of frequent glucose monitoring, another
activity which is required of a diabetes patient for optimal
management of the disease, which is performed for example as often
as five times daily.
[0102] Remission from diabetes due to successful islet neogenesis
therapy in combination with an immunosuppressive agent is indicated
by a decreased fasting blood level of glucose, and by a decreased
level and duration of elevated blood glucose in response to a
dietary challenge of sugar consumption. Upon achieving successful
islet neogenesis, insulin administration is reduced from, for
example, five injections to two injections per day; from two
injections to one injection per day; and from one to none, as
indicated by data obtained from monitoring blood glucose levels.
One of ordinary skill in the art of diabetology, when treating a
diabetic patient, is familiar with adjusting insulin dosage to
levels of blood glucose following fasting and under other
physiological conditions.
[0103] Dosages of the compositions to be administered to a subject
are adjusted for known variations from species to species in
standard data encompassing criteria for absorption, distribution,
half-life kinetics in circulation, metabolism, excretion, and
toxicology of the receptor ligands of the embodiments herein, for
example, for each primate and rodent species. In general, dosages
are adjusted to be about 6-fold to about 100-fold greater for
administration to a rodent species than to a primate species.
[0104] Immunosuppressive agents in Table 1 or other equivalent
agents are administered as supplied by the manufacturers,
normalizing to body weight of the subject as is known by one of
skill in the pharmacological arts. For example, Tacrolimus is
generally administered by injection or orally, and Sirolimus is
generally administered orally.
[0105] Modes of administration of the receptor ligand compositions
and immunosuppressive agents include but are not limited to
subcutaneous, transdermal, intrathecal, intramuscular,
intraperitoneal, intravenous, intranasal, and oral routes. The
compounds may be administered by any convenient route, for example,
by infusion or bolus injection, by pump, by absorption through
epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and
intestinal mucosa, etc.). The receptor ligands herein may be
administered in combination with one or a plurality of other
biologically active agents. For example, a recipient of the
compositions and methods herein may be administered one or more
antibiotics if a bacterial infection is present, or aspirin if a
headache is present. Administration of the receptor ligands herein
is preferably by a systemic route.
[0106] The invention in one embodiment provides a method for
treating diabetes mellitus by administering a composition
comprising both a gastrin/CCK receptor ligand, e.g. gastrin, and an
EGF receptor ligand, or a FACGINT, e.g. GLP-1, GH, or prolactin in
a formulation for sustained release, to effect differentiation of
pancreatic islet precursor cells to mature insulin-secreting cells.
Both the FACGINT and the gastrin/CCK receptor ligand in the
composition can be administered systemically or locally.
[0107] Without being limited by any particular mechanism, prolonged
efficacious islet cell neogenesis is achieved following
administration of both a gastrin/CCK receptor ligand, such as
gastrin, and an EGF receptor ligand or one or more of a member of
the FACGINT group as defined herein, such as a GLP-1, a GH, or a
prolactin, alone or with an agent for immune suppresion, and any
one or more of these receptor ligands or factors or agents being
formulated for sustained release as described herein or by
equivalent methods known or one of ordinary skill in the
pharmacological arts.
[0108] The invention in a general embodiment provides a method for
preventing or treating diabetes, the method comprising
administering to a mammal in need thereof a composition comprising
a sustained release formulation of at least one of an EGF receptor
ligand or a FACGINT, such as a GLP-1, an exending-4, a growth
hormone, a prolactin, in combination with a gastrin/CCK receptor
ligand, each of the EGF receptor ligand or the FACGINT and the
gastrin/CCK receptor ligand in an amount sufficient to increase the
number of pancreatic insulin secreting .beta. cells in the mammal,
thereby treating or preventing the diabetes. Compositions for
treating diabetic subjects and patients with a sustained release
formulation of a FACGINT comprise : a PTH-related protein (PTHrP)
receptor ligand such as PTHrP (PTHrP; Garcia-Ocana, A. et al.,
2001, J. clin. Endocrin. Metab. 86: 984-988); a hepatocyte growth
factor (HGF) receptor ligand such as HGF (HGF; Nielsen, J. et al.,
1999, J Mol Med 77: 62-66); a fibroblast growth factor (FGF) such
as FGF, a keratinocyte growth factor (KGF) receptor ligand such as
KGF; a nerve growth factor (NGF) receptor ligand such as NGF; a
gastric inhibitory polypeptide (GIP) receptor such as GIP; a
transforming growth factor beta (TGF.beta.) receptor ligand such as
TGF.beta. (U.S. patent application No. 2002/0072115 published Jun.
13, 2002), a laminin receptor ligand such as laminin-1; an islet
neogenesis associated protein (INGAP) receptor ligand such as
INGAP; a bone morphogenetic factor (BMP) receptor ligand such as
BMP-2; a vasoactive intestinal peptide (VIP) receptor ligand such
as VIP; a glucagon-like peptide 1 receptor ligand such as GLP-1 and
exendin-4, glucagon-like peptide 2 (GLP-2) receptor ligand such as
GLP-2, and dipeptidyl peptidase IV inhibitors which indirectly
affect the levels of GLP-1 (Hughes, T. et al., 2002, Am Diabetes
Assoc Abstract 272-or) by inhibiting an enzyme involved in its
integrity; a REG receptor ligand such as REG protein; a Growth
hormone (GH) receptor ligand such a GH, a Prolactin (PRL) receptor
ligand such as PRL and placental lactogen (PL); an Insulin-like
growth factor (Type 1 and 2) receptor ligands such as IGF1 and
IGF-2; an Erythropoietin (EPO) receptor ligand such as EPO
(http://www.drinet.org/html/august.sub.--2002_.htm); a betacellulin
(also considered to be a member of the EGF family); an Activin-A
receptor ligand such as Activin-A; a vascular Activin-A; a vascular
endothelial growth factor (VEGF) receptor ligand such as VEGF; a
bone morphogenesis factor (BMP) receptor ligand such as BMP-2; a
vasoactive intestinal peptide (VIP) receptor ligand such as VIP; a
vascular endothelial growth factor (VEGF) receptor ligand such as
VEGF; a pituitary adenylate cyclase activating polypeptide (PACAP)
receptor ligand such as PACAP; a granulocyte colony stimulating
factor (G-CSF) receptor ligand such as G-CSF; a
granulocyte-macrophage colony stimulating factor (GM-CSF) receptor
ligand such as GM-CSH; a platelet-derived growth factor (PDGF)
receptor ligand such as PDGF and a Secretin receptor ligand such as
secretin. These sustained release formulations may also include an
agent for immune suppression.
[0109] In one embodiment, the sustained release formulation of the
composition is administered systemically. Alternatively, the
composition is administered locally, or with a device or means for
local delivery. The mammal is a diabetic mammal, for example, the
mammal has been diabetic for an extent of at least about 1% of the
lifespan of the mammal. In general, the amount of the sustained
release formulation of the gastrin/CCK receptor ligand, the FACGINT
or the EGF receptor ligand in the composition is substantially
lower than the minimum effective dose of any of these alone
required to reduce blood glucose in the diabetic mammal. The
FACGINT or the EGF receptor ligand and the gastrin/CCK receptor
ligand are provided in amounts sufficient in combination to induce
differentiation of the pancreatic islet precursor cells into
glucose responsive insulin secreting islet cells for a prolonged
period of time.
[0110] Another embodiment of the invention provides a method for
preventing or treating diabetes, the method comprising
administering to a mammal in need thereof a sustained release
formulation of a composition comprising a combination of a FACGINT
or an EGF receptor ligand, and a gastrin/CCK receptor ligand, in an
amount sufficient to increase proliferation of islet precursor
cells in pancreatic tissue, thereby treating or preventing the
diabetes.
[0111] In another aspect, the invention provides a method for
preventing or treating diabetes, the method comprising
administering to a mammal in need thereof a sustained release
formulation of a composition comprising a combination of a FACGINT
or an EGF receptor ligand, and a gastrin/CCK receptor ligand, each
in an amount sufficient to increase the number of pancreatic
insulin secreting .beta. cells in the mammal; and determining the
amount of islet neogenesis, thereby treating or preventing the
diabetes. Administering the composition reduces blood glucose
compared to blood glucose assayed prior to administering the
composition, for example, administering the composition reduces
blood glucose by about 50%, or by about 70%, compared to blood
glucose assayed prior to administering the composition.
Glycosylated hemoglobin concentration is reduced compared to
glycosylated hemoglobin concentration in the mammal assayed prior
to administering the composition. Serum insulin concentration is
increased compared to serum insulin concentration in the mammal
assayed prior to administering the composition. Pancreatic insulin
concentration is increased compared to pancreatic insulin
concentration in the mammal assayed prior to administering the
composition.
[0112] In another aspect, the invention provides a method for
inducing pancreatic islet neogenesis in a mammal, the method
comprising administering to the mammal a sustained release
formulation of a composition comprising a combination of a FACGINT
or an EGF receptor ligand and a gastrin/CCK receptor ligand, each
in an amount sufficient to increase proliferation of islet
precursor cells in pancreatic tissue, thereby inducing pancreatic
islet neogenesis.
[0113] In another aspect, the invention provides a method for
inducing pancreatic islet neogenesis in a mammal, the method
comprising administering a composition comprising a sustained
release formulation of a combination of a FACGINT or an EGF
receptor ligand and a gastrin/CCK receptor ligand, each in an
amount sufficient to increase the number of pancreatic insulin
secreting .beta. cells in the mammal.
[0114] In another aspect, the invention features a composition
comprising a gastrin/CCK receptor ligand, and a FACGINT or an EGF
receptor ligand, any of which agents are formulated in sustained
release formulation. The composition is in a dosage effective for
inducing proliferation of islet precursor cells into an increased
amount of mature insulin secreting cells. Further, the composition
is in a dosage effective for inducing differentiation of an islet
precursor cell into a mature insulin secreting cell. The
composition can be in a pharmaceutically acceptable carrier. The
composition can include an agent for suppression of an immune
response.
[0115] In another aspect, the invention provides a kit for treating
or preventing diabetes, containing a composition comprising a
gastrin/CCK receptor ligand and a FACGINT or an EGF receptor
ligand, any of which are present in sustained release formulation,
a container, and instructions for use. The composition can include
an agent for immune suppression. The composition of the kit can
further comprise a pharmaceutically acceptable carrier. The
composition of the kit can be present in a unit dosage.
[0116] Another embodiment of the invention provided herein is a
method for expanding and differentiating stem cells in a diabetic
recipient of the cells into insulin secreting cells, comprising
implanting the cells in the recipient, and administering a
composition containing an effective dose of each of a gastrin/CCK
receptor ligand and a FACGINT or an EGF receptor ligand, one or
more of which are present as a sustained release formulation. For
example, the implanted cells are obtained from a human. Further,
the implanted cells are obtained from pancreatic islets, umbilical
chords, embryos, or stem cell lines. Generally, the gastrin/CCK
receptor ligand is human gastrin 1-17Leu15. Generally, the EGF
receptor ligand is an EGF or a TGF.alpha., or is a polypeptide that
structurally is substantially identical to EGF or TGF.alpha.,
respectively, and has substantially the same biological functions
of each of EGF or TGF.alpha.. In a related embodiment, cells, for
example, stem cells are implanted by a route selected from: direct
injection into an organ, and by intravenous administration. For
example, the cells are injected into an organ selected from the
pancreas, the kidney, and the liver. Alternatively, the cells are
administered to the portal vein using a percutaneous or
transhepatic route. In either case, prior to implanting, the cells
can be treated ex vivo with the composition.
[0117] Another embodiment of the invention provided herein is a
method of treating human diabetes by implanting a reduced amount of
stem cells into a diabetic recipient, the method comprising
administering to the recipient an effective dose of each of a
sustained release formulation of a gastrin/CCK receptor ligand and
a FACGINT or an EGF receptor ligand, the amount of cells reduced in
comparison to implanting cells into an otherwise identical
recipient in the absence of administering the effective dose. The
recipient can further be administered an agent that suppresses the
immune system. For example, the agent is a chemical selected from
the group consisting of FK506, rapamycin, cyclosporin and
cortisone. Alternatively, the agent is an antibody, for example,
the antibody is anti-CD4. In any of the methods provided that
involve stem cells, the cells prior to implanting can be obtained
from a family member and stored for later use.
[0118] An embodiment of the present invention provides improved
methods and compositions for use of a FACGINT or an EGF receptor
ligand administered with a gastrin/CCK ligand, any one or more of
these agents being formulated for sustained release, to treat
diabetes. The present invention in one embodiment provides a
sustained release formulation of any of: a gastrin, in combination
with a FACGINT or an EGF receptor ligand, to achieve greater
efficacy, potency, and utility than achieved with a FACGINT or an
EGF receptor ligand alone, or with a combination of any of these
agents administered to provide immediate bioavailability of the
entire formulation. The present invention provides an improved
therapeutic ratio for the sustained release formulation compared to
the immediately bioavailable formulation.
[0119] Use of a sustained release formulation can extend the
reduction in blood sugar for an even greater period of time.
[0120] The combination of a gastrin/CCK receptor ligand and an EGF
receptor ligand or FACGINT in a sustained release formulation, with
systemic administration, had greater potency than when it was
administered in a non-sustained release direct formulation. An
improvement in glucose tolerance and an increase in pancreatic
insulin levels were observed with sustained release formulation of
the gastrin/FACGINT combination or gastrin/EGF receptor ligand
combination.
[0121] The method for treating diabetes mellitus in an individual
in need thereof includes administering to the individual a
sustained release formulation of a composition that provides both a
gastrin/CCK receptor ligand and a FACGINT, or gastrin/CCK receptor
ligand and an EGF receptor ligand, the compositions in doses
sufficient to effect differentiation of pancreatic islet precursor
cells to mature insulin-secreting cells. The cells differentiated
are residual latent islet precursor cells in the pancreatic duct. A
method for treating insulin dependent diabetes, especially Type I
or juvenile diabetes mellitus, comprises administering, preferably
systemically, a differentiation regenerative amount of both a
gastrin/CCK receptor ligand and a FACGINT or a gastrin/CCK receptor
ligand and an EGF receptor ligand, either or both agents in a
sustained release formulation, to a diabetic mammal, to stimulate
islet neogenesis to increase the number of functional glucose
responsive insulin secreting .beta. cells in the pancreas. The
combination of a gastrin and either a FACGINT or an EGF receptor
ligand, any of which in a sustained release formulation, would
result in a significant enhancement of the islet neogenesis
response over that observed with the same agents but in a
non-sustained release direct formulation. An exemplary gastrin/CCK
receptor ligand is gastrin or its synthetic gastrin derivative as
described herein, and exemplary FACGINTs are GLP-1, GH, and
prolactin. An exemplary EGF receptor ligand is a recombinant human
EGF, for example, EGF51N, a 51 amino acid long human recombinant
mutant EGF having a deletion at the C-terminal and an asparagine
residue at position 51.
[0122] Alternatively, the cells are administered to the portal vein
using a percutaneous or transhepatic route. In either case, prior
to implanting, the cells can be treated ex vivo with the
composition.
[0123] Formulations for Sustained Release and Local Delivery and
Methods of Administration
[0124] Administration of at least one of the agents that are a
gastrin/CCK receptor ligand, or an EGF receptor ligand or a
FACGINT, is formulated to occur by sustained or controlled release.
"Sustained release" as used here and in the claims refers to a
combination of materials, devices, formulation, and/or
administration of at least one I.N.T..TM. therapeutic agent, that
creates a continuous or discontinuous, such as cyclical, supply of
an amount of the combination or at least one I.N.T. agent, or
immune suppressing agent, to the recipient as a function of time.
The period of time over which release can be sustained is minutes,
hours, days, weeks, or even months. The release of the active agent
may be constant over a long period, alternatively release may be
cyclical over the prolonged period release. Release may be
independent of conditions, alternatively release may be triggered
in response to a composition in the environment or to other
external events. For example, release may be triggered by an
endogenous composition such as insulin or glucose, or release may
be triggered by an externally supplied composition such as in
response to administration of a drug.
[0125] Sustained release is contrasted to a non-sustained release
formulation that delivers the whole of the therapeutic agent dose
instantaneously or very rapidly, or is bioavailable in the entirety
of the composition in a very short period following administration,
making a large proportion of the agent available to the recipient
over a very short time period. Examples of substantially
instantaneous bioavailability of a dose include aqueous solutions
of agents that are administered parenterally, for example by a
bolus intraperitoneal injection, or orally, or even administered by
intravenous drip over a period of several hours. Thus, intravenous
administration of a cardiovascular imaging agent is circulated
throughout the arterial system within 15 seconds; intravenous drip
administration of an anti-tumor agent is entirely bioavailable
within seconds of finishing the drip. In contrast, sustained
release formulations benefit the recipient patient both by
providing long term bioavailability, and by protein the patient by
avoiding potential side effects that might result from initial high
levels of the agent. See Mathiowitz, E., Encyclopedia of Controlled
Drug Delivery. 1999. New York: John Wiley.
[0126] Sustained release formulations have been developed to
provide systemic as well as local (or targeted) administration of
agents. A variety of materials, devices and routes of
administration reviewed herein is used with the I.N.T..TM. agent
such as the gastrin/CCK receptor ligand, and the EGF receptor
ligand or the FACGINT.
[0127] Oral sustained release systems have been available, for
example, hard gelatin capsules containing a plurality of types of
pellets, each type having a different thickness of coating such
that some pellets are uncoated more quickly. See Banga, A., Bus.
Brief.: Pharmatech 2002: 151-154. In oral osmotic systems, fluid
entering a tablet through a semi-permeable membrane generates an
osmotic pressure of core components that is independent of
variables in the gastrointestinal tract. To protect agents that are
macromolecules, for example proteins such as growth factors,
methods include use of site-specific delivery, protease inhibitors,
carrier systems, or formulations such as hydrogels that contain
polyacrylic acid backbones and bioadhesive properties.
[0128] Most protein agents are delivered parenterally, and
controlled release methods for parenteral delivery of proteins
include use of lactic acid-based polymers such as poly(D,
L-lactide-co-glycolide; PLGA), which forms biodegradable
microspheres with a core containing the agent, and that release the
agent over a course of time of about one month. Further, proteins
or liposomes (see below) containing protein can be pegylated by
covalent addition of polyetheylene glycol (PEG).
[0129] Transdermal delivery can be used both for systemic and for
local delivery. Permeation enhancers as described infra can be used
in combination with a transdermal patch to improve delivery, or in
combination with a device for iontophoresis, phonophoresis,
microporation, or elecroporation with possible wearable electrical
devices.
[0130] The term, "local delivery" as used herein refers to
administering by a route and formulation or device or both such
that a particular target organ or tissue is substantially treated
while other organs and tissues are not so treated, or that the
extent of treatment of the target tissue or organ receives at least
two-fold, at least five-fold, or at least 10-fold greater dose than
non-target tissues or organs. In general herein, a target organ is
the pancreas. As shown herein, cells for transplantation or
multi-cellular transplants can be locally delivered to the pancreas
by injection into the portal vein; drugs can be delivered by
injection into pancreatic arteries, hepatic arteries, portal vein,
or pancreatic duct. It is possible to employ a pump that is not
implanted, i.e., remains external and delivers the composition to a
target organ such as the pancreas by a catheter connecting the pump
to the organ. It is also possible to employ an implantable pump to
deliver the composition locally.
[0131] Other procedures for local delivery include without
limitation endoscopic retrograde cholangiopancreatography (ERCP);
endoscopic ultrasound-guided fine needle aspiration biopsy
(EUS-FNAB) which is adapted for delivery of pharmaceutical
compositions provided herein, rather than used for sampling. See
for example, Wang et al., Transpl. Int 1995, 8: 268-272. While
these technologies are devised for diagnostic or prognostic
purposes, they can be adapted for delivery of compositions herein,
which may I.N.T..TM. compositions as described herein, to be
combined with immune suppressing agents, and or as sustained
release formulations. See also, Yano et al., 1994, Transpl Int 7
Suppl 1:S187-193; Ricordi et al., 1994, Transpl Proc 26: 3479; and
Munoz-Acedo et al., 1995, J Endocrin 145: 227-234.
[0132] Pumps which can be implantable or non-implantable (external)
pumps for drug delivery are in use for treatment of several
diseases, for example, for cancer and for diabetes. A pump can be a
peristaltic pump, a fluorocarbon propellant pump, or an osmotic
pump including a mini-osmotic pump (Blanchard, S., 1996 Biomedical
Engineering Applications, North Carolina State University).
Peristaltic pumps deliver a set amount of drug with each electric
pulse that drives the pump head. The pump, electronics and power
source are located in a titanium housing covered in Silastic. Drug
reservoirs are silicone rubber pouches that can withstand
substantial pressure, for example, 60 psi. The reservoir can be
refilled percutaneously through a polypropylene port. Fluorocarbon
pumps use a fluorocarbon liquid to operate the pump. Osmotic pumps
use osmotic pressure to release the drug at a constant rate. An
exemplary pump is the MiniMed MicroMed 407C pump (Medtronic, Inc.,
Northridge, Calif.). Further, an intrathecal drug delivery system
(Medronic) which includes two implantable components, an infusion
pump, and an intraspinal catheter, can be used. The pump is
inserted abdominally in a subcutaneous pocket, while the catheter
is inserted into the intrathecal space of the spine, tunneled under
the skin, and connected to the pump. Medication is then delivered
at a constant or variable flow rate. Further, an intraperitoneal
pump, for instance an implantable pump, can be used to deliver the
compositions herein locally, for example via a catheter, or
systemically.
[0133] Mucosal delivery to epithelial areas that are maintained in
a moist condition and which have close underlying vasculature can
be more efficient than transdermal delivery across a tissue which
is a dry epidermal surface. Mucosal surfaces include: nasal,
pulmonary, rectal, buccal, ocular, and genital. Exemplary mucosal
surfaces are nasal and pulmonary.
[0134] Materials used for microspheres for sustained release are
primarily polymers, and include PEG, also called polyethylene oxide
(PEO), and PGLA as described. A polymer vehicle can be injected
directly, allowing for gradual hydrolysis or degradation within the
subject to release the therapeutic agent. The molecular weight of
the polymer, e.g., PEG, can be varied to control the release rate.
Pegylation or covalent attachment of PEG to a protein therapeutic
agent shields the protein from receptors involved in clearance
mechanisms such as that of the reticulo-endothelial system (RES).
Alternatively, polysaccharides can be used to target an agent to
the RES (U.S. Pat. No. 5, 554,386 issued Sep. 10, 1996). Organs of
the RES include liver, spleen, and bone marrow.
[0135] Homopolymers or co-polymers, such as poly(lactic
acid-co-ethylene glycol) or PLA-PEG, can form a viscous liquid
which is mixed with a therapeutic protein agent. Viscosity is
varied by the molecular weight of the PLA-PEG. Under certain
conditions the therapeutic agent co-precipitates with the polymer
upon injection into the subject and loss of the solvent by
diffusion, such that a depot is formed having favorable release
kinetics (Whitaker, M., et al. Bus. Brief: Pharmatech
2002:1-5).
[0136] Microparticles are formed of polymeric microspheres that
encapsulate the therapeutic agent. Polymers for use in microspheres
include poly(lactic acid) or PLA; poly(glycolic acid) or PGA; and
copolymer PLA-PGA. An amount of the therapeutic agent such as the
proteins of the present invention, is released in stages such as an
initial burst of protein non-specifically associated with the
exterior of the particles, a later stage by diffusion, and a final
stage by erosion can be controlled by polymer composition,
molecular weight, size of the microparticles, and physiological
conditions such as pH. Protein stability during manufacture of
microspheres is enhanced if the protein is in solid form, such as a
lyophilized powder, which is emulsified with solvent or by a
frozen-atomization process or a sonication process, and the
suspension is then frozen in liquid nitrogen for further solvent
extraction. Microspheres can be produced from a supercritical
fluid, e.g., supercritical carbon dioxide (scCO.sub.2).
[0137] Biodegradable block polymers that are suitable for drug
delivery and methods of synthesis are described by Kumar, N. et
al., Adv. Drug Deliv. Rev. 53 (2001): 23-44. Copolymers can be
random, alternating, or block (di or tri type) and can be linear,
or star or graft (comb-shaped) in configuration. A polymer can form
a hydrogel, which is a three-dimensional, hydrophilic polymeric
network that holds a large amount of aqueous fluid. The polymer
used in a hydrogel is rendered insoluble by cross-linking or other
chemical adducts.
[0138] Biodegradable implants can be prepared from materials such
as at least one of the materials selected from the group of:
starch; vinylstarch; dipropyleneglycol diacrylate (DPGDA);
tripropyleneglycol diacrylate (TPGDA); pectin; cellulose acetate;
cellulose propionate; cellulose acetate butyrate; cellulose acetate
propionate (CAP); hydroxypropyl cellulose (HPC); hydroxypropyl
cellulose/cellulose acetate propionate (HPC/CAP); methyl
methacrylate (MMA); butyl methacrylate (BMA); hydroxymethyl
methacrylate (HEMA); ethyl hexyl acrylate (EHA); octadecyl
methacrylate (ODMA); and ethyleneglycol dimethacrylate (EGDMA). See
Gil, M. et al., Boletim de Biotecnologia 2002, 72:13-19.
[0139] In addition to polymers, naturally occurring and synthetic
lipids can be used for sustained release formulations. DepoFoam.TM.
(Skye Pharma, London, England) forms a multivesicular lipid-based
particle (liposome) for encapsulating therapeutic agents (see U.S.
Pat. No. 5,993,850; and Ye, Q. et al., 2000 J. Controlled Rel.
64:155-166). The lipids are amphipathic with a net negative charge,
sterols, or zwitterionic lipids, and methods for making the
liposomes are non-acidic. Methods for incorporating an active
therapeutic agent into the liposomes are also provided. The
therapeutic agent can be one or more of a gastrin/CCK receptor
ligand, an EGF receptor ligand, and a FACGINT.
[0140] Other lipids for liposomes are within the scope of the
invention. A plant polar lipid, for example a ceramide such as a
wheat ceramide, is useful for forming a gel with a protein such as
a prolamine, into which one or more therapeutic agents can be
placed for transdermal or transmucosal delivery. See U.S. Pat. No.
6,410,048, issued Jun. 25, 2002. Exemplary prolamines include wheat
gliadin, and corn zein. Other naturally occurring polymers used in
sustained release drug formulations and devices include collagen
(EP-A-O 621 044), chitin (U.S. Pat. No. 4,393,373), and chitosan, a
deacylated form of chitin.
[0141] A permeation enhancer, for example, a glycolipid, a
non-esterified fatty acid, an aliphatic alcohol, a fatty acid ester
of an aliphatic alcohol, a cyclohexanol, a fatty acid, ester of
glycerol, a glycol, or an aliphatic alcohol ether or a glycol, are
typical permeation enhancers, other components such as a
stabilizer, a solubilizer, a surfactant and a plasticizer can be
present in a transdermal device. See U.S. patent application No.
20020127254, published Sep. 12, 2002.
[0142] Lipids and a variety of types of polymers are used to form
"nanoparticles" for drug delivery, reviewed by M. Kumar, 2002, J.
Pharm. Pharmaceut. Sci. 3:234-258. Drug loading into these
particles is found to be greatest with most lipophilic therapeutic
agents. Long lasting release has been found with liposomes
comprising polylactic acid, lecithin, and phosphatidylcholine or
cholesterol.
[0143] Local (targeted) sustained release is obtained by methods
described herein, for example, use of liposomes comprised of a
material designed to target the RES, or comprised of a different
material to avoid cells of the RES. Additional targeting methods
include use of antibodies or soluble recombinant receptors in
conjunction with the outer surface of liposomes or microspheres.
Further, sustained release formulations as described herein may be
further adjusted for use with any of the devices described herein,
such as a pump, for local delivery to a particular target
organ.
[0144] The present invention in another embodiment provides
sustained release pharmaceutical compositions comprising a
therapeutically effective amount of a combination of a FACGINT or
an EGF receptor ligand, and a gastrin/CCK receptor ligand. A
pharmaceutically acceptable carrier or excipient can be added. Such
a carrier includes but is not limited to saline, buffered saline,
dextrose, water, glycerol, ethanol, and combinations thereof. The
formulation should suit the mode of administration.
[0145] Unless otherwise defined, all technical and scientific terms
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention pertains. Methods
and materials similar or equivalent to those described herein can
be used in the practice of the present invention. The invention in
various embodiments now having been fully described, additional
embodiments are exemplified by the following Examples and claims,
which are not intended to be construed as further limiting. The
contents of all cited references are hereby incorporated by
reference in their entirety herein.
EXAMPLES
Example 1
Treatment With a GLP-1 Receptor Ligand, Glucagons-Like Peptide 1
(GLP-1) and a Gastrin/CCK Receptor Ligand, Gastrin, Prevents
Disease Progression in NOD Mice With Recent-Onset Diabetes.
[0146] Mice of the non-obese diabetic (NOD) strain have a phenotype
that shares many features of disease pathogenesis with human type I
diabetes. NOD mice typically exhibit destructive autoimmune
pancreatic insulitis and .beta.-cell destruction as early as four
weeks of age. Diabetes onset usually occurs at age 10-15 weeks in
these mice, with typical blood glucose levels observed to be
between about 7 mM to about 10 mM (compared to a range of about
3.0-6.6 mM normal mice), and a pancreatic insulin level that is
lower by more than about 95% than that in normal mice. As disease
progresses, NOD mice exhibit increasingly severe signs of chronic
diabetes, with blood glucose levels reaching between about 25 to
about 30 mM, and pancreatic insulin level declining to become
virtually non-existent. At that severe stage of the disease,
greater than about 99% of .beta.-cells have been destroyed.
[0147] In this example, the effect of treatment by a combination of
GLP-1 and gastrin was examined in NOD mice with recent onset
diabetes, to determine whether administration of both GLP-1 and
gastrin would prevent severe hyperglycemia, ketoacidosis and death
as well as increase pancreatic insulin content in NOD mice with
recent-onset diabetes. The I.N.T..TM. composition used was gastrin
as synthetic human gastrin I having 17 amino acid residues with a
Leu residue at amino acid position 15. The GLP-1 used was GLP-1
which is the biologically active fragment of human/mouse GLP-1
(having residues at positions 7-36 compared to the precursor from
which the fragment is processed; obtained from Bachem H6795).
[0148] Non-obese diabetic (NOD) female mice, ages 12-14 weeks, were
monitored for development of onset of diabetes (fasting blood
glucose>8.0 to 15 mmol/l), and within 48 hours after onset of
symptoms, two groups of mice were each treated as follows: one
group was treated with vehicle only; and the other group was
administered 100 .mu.g/kg/day of GLP-1, each treatment administered
via the intraperitoneal route twice daily.
[0149] Therapy was administered for 14 days. Animals were monitored
weekly for fasting blood glucose (FBG) levels. FBG levels were
measured at about 12 hours after food had been withdrawn, and 24
hours after the last peptide or vehicle injection. Upon cessation
of therapy, all mice were monitored for FBG levels for the next 4
weeks (weeks 2-6) so as to determine whether prevention of
hyperglycemia persisted after termination of therapeutic treatment.
At 14 days treatment was stopped, and at 18 days the mice were
sampled to obtain FBG levels (as shown in Table 2).
2TABLE 2 Glucagon-like peptide - 1 (GLP-1) and gastrin combination
therapy treats recent onset diabetes in NOD mice FBG (mM) at day:
Group GLP-1 Gastrin Number 0 7 14 18 1 - - 6 11.0 .+-. 0.6 14.8
.+-. 1.3 22.8 .+-. 1.6 24.4 .+-. 1.5 2 + - 4 12.3 .+-. 0.9 14.1
.+-. 1.8 15.3 .+-. 2.6 15.8 .+-. 4.2 3 - + 4 11.8 .+-. 0.9 14.8
.+-. 3.4 16.4 .+-. 3.4 19.0 .+-. 4.5 4 + + 4 13.5 .+-. 0.9 10.4
.+-. 0.4 7.9 .+-. 0.8 7.9 .+-. 1.5 GLP-1 (100 .mu.g/kg/day) and
gastrin (3 .mu.g/kg/day) were administered by i.p. injection to NOD
female mice, twice a day, to each group as shown above. Diabetic
mice, aged 12-14 weeks, were used within two days of diabetes onset
(generally considered to be an FBG of more than 6.5 mM).
[0150] The protocol includes sampling of these mice for data again
at 6 weeks, and blood collected for assay of FBG and plasma
C-peptide, and the mice are sacrificed for pancreatic insulin
determinations and scoring of islet inflammation (insulitis). From
the outset of treatment, mice received neither insulin-replacement
treatment nor immunosuppression. The following parameters are
assessed: survival rates, pancreatic insulin levels, presence of
islet inflammation and fasting blood glucose levels.
[0151] Results show that in animals of the vehicle-treated control
group (group 1), fasting blood glucose (FBG) values increased
progressively during the time course of this mock treatment, from
11 mM glucose at day 0 to 24 mM at day 18.
[0152] In contrast, in mice in treated with GLP-1 and gastrin, FBG
values (7.9 mM glucose) were significantly less compared to the
vehicle-treated mice (24.4 mM), reduced to a level, in fact, that
is less than one third of the level that developed in the vehicle
trated mice; Table 2). Most significant and surprisingly, the
combination of GLP-1 and gastrin was more effective in decreasing
FBG levels than either gastrin alone or GLP-1 alone (having FBG
levles of 19.0 mM and 15.8 mM, respectively). Only in mice treated
with the combination was the FBG reduced to a level that is in a
normal range. The improved control of blood glucose levels in mice
treated with GLP-1 and gastrin may be expected to be associated
with a significantly increased content of insulin in the pancreas
of these mice.
[0153] In summary, the results in this study show that treatment
with a short course of low doses of GLP-1 and gastrin treatment in
mice with recent onset of diabetes prevented disease progression,
and reversed the disease condition to yield a blood glucose level
of about normal. Further, this significant decreased blood glucose
level in mice treated with gastrin and GLP-1 persisted for an
additional time period after cessation of treatment. It is expected
that in these animals, data will show improved pancreatic insulin
content, and that these effects will be shown to be sustained for a
prolonged period of time after termination of therapy.
[0154] Further groups of NOD female mice were treated with a
prolactin receptor ligand, prolactin (PRL), and a gastrin/CCK
receptor ligand, and with a growth hormone receptor ligand, growth
hormone (GH), and a gastrin/CCK receptor ligand, gastrin. It is
anticipated that data will indicate that these treatments yield
effects on FBG and other parameters that are similar to data
obtained herein with GLP-1.
Example 2
Comparison of Compositions With and Without Pegylation
[0155] In order to determine whether a sustained release
formulation would provide greater efficacy than an otherwise
identical formulation that is formulated for bolus or non-sustained
release delivery, a study is performd comparing a protocol of
treating NOD mice with an I.N.T..TM. composition either in a
pegylated or non-pegylated formulation. Pegylation of at least one
component of the I.N.T..TM. composition can prolong the amount of
time the therapeutic agent is retained ina na active form in the
body.
[0156] The study shows that administration of a sustained release
formulation of at least one agent of an I.N.T..TM. composition is
able to improve the diabetic conditions of the NOD mice, as
compared to an I.N.T..TM. composition formulated for direct, i.e.,
non-sustained release administration.
Example 3
Comparison of Frequency of Dosages Composition Administration
[0157] In order to determine whether a sustained release
formulation would provide greater efficacy than a non-sustained
release direct formulation, an experiment is devised having
comparing a protocol of three-time a day administration to the
once-a-day administration of an I.N.T..TM. composition to NOD
mice.
[0158] The results show that prolongation of bioavailability of the
I.N.T..TM. composition, comparable to that obtained with a
sustained release formulation of an I.N.T composition, can improve
efficacy, i.e., can better remediate symptoms of the diabetic
condition of the NOD mice, as compared to a direct or non-sustained
release formulation of an I.N.T..TM. composition, and can reduce
the frequency of dosages required for such remediation of
symptoms.
* * * * *
References